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Poste Italiane spa - Spedizione in A.P. - D.L. 353/2003 (conv. in L. 27/02/2004 N. 46) art. 1, comma 1, DCB Piacenza - Concessionaria esclusiva per la pubblicità: E.V. Soc. Cons. a r.l. - Cremona
LAR
LARGE ANIMAL REVIEW è indicizzata su Science Citation Index (SciSearch®)
Journal Citation Reports/Science Edition e CAB ABSTRACTS
Bimestrale, Anno 23, Numero 5, Ottobre 2017
Large Animal Review
05/17
ARTICOLI ORIGINALI
BOVINI
• Stima dell’effetto pascolo sul profilo
in acidi grassi del latte bovino
in allevamenti di montagna
• Bovine papillomatosis in Sicily:
virus identification, diagnosis and
co-infections
OVINI
• Interbreed variation in craniometrical
parameters in sheep
• Monitoring of selected milk quality
parameters during lactation of a
Mediterranean dairy sheep breed
CAPRINI
• Preliminary genetic variability analysis
of the native Garfagnina goats based
on microsatellite polymorphism
CASE REPORTS
BOVINI
• Autologous prosthesis for the surgery
of two simultaneous hernias in a calf
ISSN: 1124-4593
0A_Copert LAR 5_2017_ok 27/09/17 12:22 Pagina 2
0A_Copert LAR 5_2017_ok 27/09/17 12:22 Pagina 3
Anno 23, numero 5,
Ottobre 2017
Rivista indicizzata su:
CAB ABSTRACTS e GLOBAL HEALTH
IF (aggiornato al 2016): 0.315
Editor in chief
Massimo Morgante
Editorial Board 2016-2018:
Roberto Bardini - Liliana Carlomagno
Cristina Casalone - Marco Colombo
Vincenzo Cuteri - Matteo Gianesella
Paolo Moroni - Paolo Pasquali
Mariano Pauselli - Giuseppe Piccione
Fabrizio Rueca - Alessandro Zumbo
Managing Editor
Matteo Gianesella
LARGE ANIMAL REVIEW è una rivista bime-
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Poste Italiane SPA - Spedizione in A.P.
D.L. 353/2003 (Conv. in L. 27/02/2004 N. 46)
Art. 1, Comma 1, DCB Piacenza
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SOMMARIO
BOVINI
Stima dell’effetto pascolo sul profilo
in acidi grassi del latte bovino in allevamenti
di montagna
G. BOLZONI, S. BETTONI, E. BUFFOLI, C. BAIGUERA,
A. MARCOLINI, G. ZANARDI 163
Bovine papillomatosis in Sicily:
virus identification, diagnosis and co-infections
L. GALLINA, F. SAVINI, A. LAVAZZA, P. DI MARCO,
R. PULEIO, M. ROCCARO, G. PURPARI, A. GUERCIO,
A. SCAGLIARINI 171
OVINI
Interbreed variation in craniometrical
parameters in sheep
M.H. OWEN, R. DAVIES, C.E. BEARD,
A.M. OLVERA RAMIREZ, N.R. MCEWAN 177
Monitoring of selected milk quality
parameters during lactation of a Mediterranean
dairy sheep breed
T.G. BONTINIS, E.C. PAPPA, K. SOTIRAKOGLOU,
A.C. PAPPAS, E. TSIPLAKOU, G. ZERVAS 185
CAPRINI
Preliminary genetic variability analysis
of the native Garfagnina goats based
on microsatellite polymorphism
F. CECCHI, C. RUSSO, C. CANTILE, G. PREZIUSO 191
CASE REPORTS
Autologous prosthesis for the surgery
of two simultaneous hernias in a calf
F. SPADOLA, G.L. COSTA, M. MORICI, C. INTERLANDI,
B. NASTASI, M. MUSICÒ 195
RUBRICHE
UNA FINESTRA SULLE AZIENDE 198
VET-JOURNAL
A CURA DI M.G. MONZEGLIO 200
N
Aggiornamento
scientifico permanente
in medicina veterinaria
l
j
0B_Somm LAR 5_2017_ok 03/10/17 16:07 Pagina 161
0B_Somm LAR 5_2017_ok 03/10/17 16:07 Pagina 162
Garantire il benessere nell’allevamento
dell’asino per la produzione di latte:
un’analisi della legislazione
G. BOLZONI1, S. BETTONI1, E. BUFFOLI1, C. BAIGUERA1,
A. MARCOLINI1, G. ZANARDI1
1 Istituto Zooprofilattico sperimentale della Lombardia e dell’Emilia Romagna, via Bianchi 9, Brescia (I)
G. Bolzoni et al. Large Animal Review 2017; 23: 163-168 163
Stima dell’effetto pascolo sul profilo
in acidi grassi del latte bovino in allevamenti
di montagna
N
Autore per la corrispondenza:
Giuseppe Bolzoni (crn.qualita.latte@izsler.it).
RIASSUNTO
La composizione in acidi grassi del latte è stata oggetto negli ultimi decenni di numerosi studi, sia per aspetti tecnologici e or-
ganolettici dei prodotti derivati, sia per quelli nutrizionali e di salubrità. La quantità, così come la composizione lipidica del
latte, sono in parte caratteri ereditari, ma in gran parte sono determinati dall’alimentazione degli animali. Lo scopo del pre-
sente lavoro è quello di caratterizzare il contenuto in acidi grassi saturi ed insaturi del latte bovino prodotto in 37 allevamenti
tipici per alimentazione e stabulazione di un’area di montagna del nord Italia (Valle Camonica), tramite una tecnica analitica
di stima indiretta basata sulla spettroscopia in infrarosso. Attraverso l’analisi di 189 campioni di latte di massa raccolti tra la
primavera e l’estate 2015, si è potuto verificare che il periodo di pascolo influisce in misura statisticamente significativa sul rap-
porto tra acidi grassi insaturi e saturi del latte: 0,42 (DS 0,025) rapporto medio nelle stalle a fondovalle vs 0,57 (DS 0,048) rap-
porto medio delle stesse mandrie al pascolo. Il presente lavoro fornisce inoltre un quadro complessivo delle caratteristiche del
latte di montagna evidenziando alcune peculiarità indotte dal periodo di pascolo. Considerato che la tecnica analitica di scree-
ning utilizzata, seppur con un’accuratezza relativamente limitata, garantisce analisi semplici, rapide, a costo molto contenuto
ed eseguibili su elevati numeri di campioni anche per periodi prolungati, riteniamo che questo tipo di attività possa concorre-
re a supportare la produzione di latte e derivati nelle aree di montagna, valorizzandone le componenti di genuinità e salubrità.
PAROLE CHIAVE
Latte, acidi grassi, FTIR, pascolo.
INTRODUZIONE
La componente lipidica del latte bovino è notoriamente la
componente più variabile; la sua quantità dipende infatti da
numerosi fattori tra i quali i principali sono: razza, alimenta-
zione, stadio di lattazione, numero di lattazioni, condizioni
climatiche e modalità di mungitura. Ma anche la composi-
zione stessa del grasso del latte presenta al suo interno ampi
margini di variabilità, in parte collegati alla genetica degli ani-
mali1ed in gran parte all’apporto alimentare ed al successivo
metabolismo ruminale2,3. Circa il 98% del grasso del latte bo-
vino è costituito da trigliceridi (il rimanente è costituito da
fosfolipidi, mono e digliceridi, acidi grassi liberi lineari e ra-
mificati); le tre molecole di acidi grassi, legate alla base fissa di
glicerolo, costituiscono la componente variabile che influen-
za sia gli aspetti nutrizionali e di digeribilità del latte e dei
prodotti derivati, sia il comportamento tecnologico nel corso
del processo di caseificazione. La composizione in acidi gras-
si è inoltre considerata uno degli elementi fondamentali per
definire la salubrità degli alimenti: si vedano a tal proposito le
raccomandazioni FAO e WHO sull’assunzione giornaliera di
acidi grassi saturi (in particolare acido miristico e palmitico)
o sul rapporto tra alcuni acidi grassi trans ed i livelli di cole-
sterolemia4,5 o, ancora, sull’effetto dell’apporto dietetico di
acido linoleico6. I latticini sono considerati in genere una ri-
levante fonte di acidi grassi nella dieta per una parte consi-
stente della popolazione mondiale; da ciò deriva l’interesse ri-
servato allo studio della composizione in acidi grassi del latte
in relazione a differenti modalità di allevamento e alimenta-
zione delle bovine. Già segnalati, ad esempio, il significativo
effetto del pascolo sul rapporto tra acidi grassi ω6 ed ω3, sul-
la quantità di insaturi o sulla riduzione dei saturi a lunga-me-
dia catena e sulla quantità di acido linoleico coniugato
(CLA)7,8. Il presente lavoro è finalizzato alla caratterizzazione
chimica e tecnologica del latte raccolto in allevamenti bovini
tipici di un’area di montagna nel nord Italia (Valle Camoni-
ca), con particolare riferimento alla composizione in acidi
grassi, alla stima del rapporto tra la frazione satura rispetto a
quella insatura ed al confronto tra il periodo di pascolo in al-
ta quota rispetto a quello di stabulazione a fondo valle.
MATERIALI E METODI
Tramite una scheda di caratterizzazione degli allevamenti di
bovine della zona, realizzata con la collaborazione del perso-
nale addetto all’assistenza tecnica sul territorio (Gruppo
Azione Locale Valle Camonica e Val di Scalve), sono stati se-
lezionati 37 allevamenti che per ubicazione, caratteristiche e
modalità di allevamento fossero rappresentativi degli “alle-
vamenti di montagna” e quindi ben differenziabili dal tipico
allevamento intensivo predominante nell’area di pianura
della stessa regione (Lombardia). Gli allevamenti selezionati
erano, complessivamente, di piccole dimensioni (media bo-
vine in lattazione 11,8) con prevalenza di vacche razza Bru-
na Alpina (65%), con quote minoritarie di Pezzata Rossa
Bolzoni_imp_ok 03/10/17 16:08 Pagina 163
– Carica batterica totale con tecnica optofluorometrica con
cella di flusso (Bactoscan FC, Foss, DK);
– Cellule somatiche con tecnica optofluorometrica con cel-
la di flusso (Fossomatic FC, Foss, DK);
– Acidità titolabile, secondo la metodica Soxhlet-Henkel.
L’analisi statistica dei risultati è stata realizzata con fogli di
calcolo Excel Microsoft.
Sono stati impiegati gli Strumenti di Analisi e/o le formule
implementate, e gli strumenti di valutazione grafica disponi-
bili nei fogli di calcolo. Le principali analisi statistiche con-
dotte hanno riguardato: la stima di statistiche parametriche
sulla base dati (media, media geometrica, deviazione stan-
dard) ed il test di significatività per la differenza fra medie di
popolazioni (test T di Student).
Nota: riteniamo importante sottolineare che la tecnica utiliz-
zata per la determinazione delle diverse frazioni di acidi gras-
si rappresenta una metodica di screening indiretto rispetto a
metodiche di riferimento in gas-cromatografia finalizzate a
quantificare i singoli acidi grassi (ISO 15885:20029); alla va-
lidazione di base fornita dal produttore deve di conseguenza
essere sovrapposta una calibrazione realizzata dal singolo la-
boratorio10 stimata dal confronto con metodiche di riferi-
mento su campioni rappresentativi del latte normalmente
analizzato. Ciò determina, che a fronte di un’elevata ripetibi-
lità, l’accuratezza della stima risulta relativamente limitata:
ripetibilità (r=2,83 *Sr) compresa tra 0,03 e 0,10 g/100 g ed
Errore Standard di stima tra 0,04 e 0,09 g/100 g11,12.
Inoltre, la porzione di UFA è ottenuta per calcolo dalla diffe-
renza tra SFA e acidi grassi totali (stimati 0,95 dei lipidi tota-
li), per cui sono possibili, per singoli campioni, incongruen-
ze tra somma matematica dei singoli valori di MUFA e PU-
FA e quello totale di UFA. D’altra parte la sempli-
cità della procedura analitica, il ridottissimo costo
e la possibilità di analizzare elevati numeri di
campioni in rapida sequenza (fino a 400/h), la
rende particolarmente idonea a questo tipo di stu-
di in cui la finalità prioritaria è il confronto tra se-
rie di campioni; la stessa metodica è stata del resto
già largamente impiegata in lavori similari negli
ultimi anni13,14.
RISULTATI E DISCUSSIONE
Caratterizzazione generale
del latte di montagna
La caratterizzazione complessiva del latte prodotto
dagli allevamenti compresi nello studio può essere
sintetizzata attraverso i valori medi e le relative de-
viazioni standard per ciascun parametro (Tabella
1). Nella medesima tabella sono riportati, come
generico elemento di confronto, i valori medi os-
servati nello stesso periodo sui campioni conferiti
al laboratorio per il sistema di pagamento del latte
in base alla qualità (QMPS) in regione Lombardia,
per un totale di circa 4.500 allevamenti in preva-
lenza di tipo intensivo e dell’area di pianura.
Il quadro risultante fornisce ovviamente soltanto
un’immagine generica del latte prodotto dagli al-
levamenti di montagna nel periodo dello studio;
seppur con notevole variabilità (testimoniata dal-
l’elevatissimo valore di deviazione standard di al-
Note:
*Valore espresso come Media Geometrica
QMPS: Sistema di pagamento del latte in base alla qualità
N.D.: non definita
164 Stima dell’effetto pascolo sul profilo in acidi grassi del latte bovino in allevamenti di montagna
(16%) e Meticcia (16%) e livello produttivo medio-basso (16
litri/capo/die), alimentazione prevalente a fieno di produzio-
ne locale con integrazione di concentrati di cereali inferiore
al 30% della razione (2-7 kg/capo/die) ed assenza di foraggi
insilati (mais o altro). Tra gli elementi fondamentali della se-
lezione degli allevamenti è stato considerato lo spostamento
degli animali in pascoli di alta quota nel periodo estivo. In
ogni allevamento, ed in ognuna delle 19 “malghe” in cui le
bovine erano trasferite per il pascolo, è stato eseguito alme-
no un campione al mese di latte di massa aziendale, sia nel
periodo di stabulazione a fondo valle (Marzo-Maggio 2015,
105 campioni “M”) che nel periodo di pascolo (Giugno-
Agosto 2015, 84 campioni “AP”). La fase di pascolo in quota
era organizzata in 19 differenti raggruppamenti di animali
(malghe): in alcune di esse accedevano anche animali prove-
nienti da più di un allevamento tra quelli controllati nella
prima fase (malghe incluse nello studio); in altri casi alla
malga accedevano invece anche animali provenienti da alle-
vamenti intensivi dell’area di pianura (malghe escluse dallo
studio). Infine, soltanto in 6 malghe si è mantenuta la per-
fetta identità di bovine presenti in 6 allevamenti di fondoval-
le del primo periodo (malghe incluse nello studio).
Su ciascun campione, addizionato di Sodio Azide come con-
servante, refrigerato fino alla consegna in laboratorio, si so-
no realizzate le seguenti determinazioni analitiche entro 48
ore dal prelievo:
– Composizione in grasso, proteine, lattosio, caseine, urea,
acidi grassi saturi (SFA), insaturi (UFA), mono e polinsa-
turi (MUFA e PUFA), con tecnica spettroscopica in medio
infrarosso con derivata di Fourier (FTIR, Milcoskan FT
6000, Foss, DK);
Grasso (g/100 g) 4,270 (0,602) 3,920
Proteine (g/100 g) 3,470 (0,229) 3,440
Caseina (g/100 ml) 2,720 (0,185) 2,680
Lattosio (g/100 ml) 5,050 (0,152) 5,160
Acidi Grassi Saturi (g/100 g) 2,570 (0,404) 2,389
Acidi Grassi Insaturi (g/100 g) 1,233 (0,272) 1,049
Acidi Grassi Monoinsaturi (g/100 g) 1,129 (0,248) 0,959
Acidi Grassi Polinsaturi (g/100 g) 0,150 (0,040) 0,123
Rapporto Insaturi/Saturi 0,470 0,430
Punto Crioscopico (°C) -0,527 (0,005) -0,524
Cloruri EQ (mg/100 ml) 150,060 (18,60) 138,000
Urea (mg/10 ml) 25,310 (7,060) 22,800
Carica Batterica Totale* (UFC/ml) 700.000 45.000
Cellule Somatiche* (cell/ml) 377.000 268.000
Acidità Titolabile (°SH/50 ml) 3,300 (0,310) N.D.
Indice caseina 0,783 (0,007) 0,777
Tabella 1
- Valori medi e deviazione standard dei parametri analitici dei 189
campioni di latte analizzati; confronto con pagamento latte qualità regionale
(QMPS).
Parametro (unità di misura) Media Campioni Media QMPS
(Deviazione Standard)
Bolzoni_imp_ok 03/10/17 16:08 Pagina 164
G. Bolzoni et al. Large Animal Review 2017; 23: 163-168 165
cuni parametri), il latte prodotto dalle diverse mandrie pre-
senta le seguenti caratteristiche principali:
1. livelli di grasso, proteine e caseine decisamente elevati,
seppur con ampia variabilità e superiori ai valori medi re-
gionali (considerato il periodo dell’anno e l’effetto della
fase di pascolo, ciò è attribuibile alla prevalenza di bovine
Bruna Alpina in montagna e la quasi esclusività della Fri-
sona in pianura);
2. contenuto in grasso ed acidi grassi del latte di montagna su-
periore a quello regionale, così come il rapporto UFA/SFA;
3. notevole differenza di contaminazione batterica che, per
quanto riguarda la montagna, è fortemente influenzata
dalle condizioni igieniche della produzione e conservazio-
ne del latte nel periodo di pascolo.
Composizione degli Acidi Grassi
nel latte di montagna
tenuto in debito conto dei limiti
Anche per quanto riguarda la composizione delle differenti
componenti acidiche, la variabilità osservata risulta consi-
stente: la distribuzione di frequenza per le due componenti
principali, UFA e SFA, per i campioni prelevati nel periodo
di stabulazione (M in grigio) rispetto a quello di pascolo (AP
in nero) è rappresentata nei Grafici 1 e 2.
Anche tramite la dispersione delle medie di allevamento
nei due periodi, è apprezzabile graficamente la tendenza
alla maggior differenziazione della componente UFA (Gra-
fico 3) rispetto a quella SFA (Grafico 4), nei due periodi di
studio.
A conferma delle osservazioni grafiche precedenti, le due se-
rie di dati sono state sottoposte ad un test statistico di signi-
ficatività delle differenze: test T (P<0,05). Il risultato ottenu-
to è sintetizzato in Tabella 2.
Tenendo in debito conto i limiti di questo tipo di inferenza,
riteniamo interessante sottolineare che:
– la componente igienico sanitaria della produzione del
latte risulta significativamente peggiorata dal passaggio
al pascolo: è facile collegare questi due aspetti alle condi-
zioni di mungitura e conservazione del latte (e dei cam-
pioni) per la carica batterica ed all’effetto del cambia-
Grafico 1
Distribuzione degli
acidi grassi insaturi nei
campioni prelevati nel
periodo di
stabulazione (M)
rispetto al periodo di
pascolo (AP).
Grafico 2
Distribuzione degli
acidi grassi saturi nei
campioni prelevati nel
periodo di
stabulazione (M)
rispetto al periodo di
pascolo (AP).
Bolzoni_imp_ok 03/10/17 16:08 Pagina 165
166 Stima dell’effetto pascolo sul profilo in acidi grassi del latte bovino in allevamenti di montagna
mento ambientale, climatico e di alimentazione per il te-
nore in cellule.
– Il valore in grasso risulta incrementato malgrado, in linea
generale, l’apporto nutrizionale al pascolo in termini di
energia sia inferiore a quello ottenibile in stabulazione. In
realtà in alcuni allevamenti dell’area di montagna l’appor-
to nutritivo nell’ultimo periodo di stabulazione cala forte-
mente per la limitata disponibilità di scorte di foraggi di
buona qualità. Va notato che l’incremento osservato si veri-
fica in un periodo dell’anno in cui negli allevamenti della
pianura padana il titolo di grasso e proteine tende tipica-
mente ad un crollo in relazione al tipico clima caldo umi-
do, evidentemente evitato con il trasferimento al pascolo.
– Risultano interessanti anche le significatività osservate per
le componenti di acidi grassi: sebbene il decremento della
componente satura risulti statisticamente non significati-
vo, le altre 3 frazioni mostrano tutte incrementi statistica-
mente significativi; il cambio di regime alimentare sembra
dunque incidere non solo sulla quantità, ma anche sulla
composizione della frazione lipidica ed in particolare sul
rapporto acidi grassi insaturi/saturi.
Considerata la possibilità di interferenze sui dati osservati
dovute, come già accennato, al parziale rimescolamento del-
le mandrie, abbiamo cercato di confermare ulteriormente le
precedenti osservazioni selezionando unicamente le 6 mal-
ghe in cui gli animali al pascolo risultavano raggruppati in
modo corrispondente alle 6 stalle di fondovalle da cui prove-
nivano (Tabella 3).
CONCLUSIONI
Le osservazioni condotte, pur con i limiti citati per quanto
riguarda la metodica analitica e la parziale variazione nella
Grafico 3
Dispersione dei valori
medi di allevamento in
acidi grassi insaturi nel
periodo di
stabulazione (M) e di
pascolo (AP).
Grafico 4
Dispersione dei valori
medi di allevamento in
acidi grassi saturi nel
periodo di
stabulazione (M) e di
pascolo (AP).
Bolzoni_imp_ok 03/10/17 16:08 Pagina 166
si tecnicamente più approfondi-
ti15,16,17 sui significativi effetti della
razione alimentare e, in particola-
re, di quella caratteristica del pa-
scolo, anche se non limitatamente
a quello di alta quota in montagna,
sulla composizione in acidi grassi
del latte. Anche con tecniche anali-
tiche indirette e decisamente sem-
plici da applicare su elevati numeri
di campioni, è possibile contribui-
re alla valorizzazione ed al soste-
gno di una filiera produttiva che,
oltre alle difficoltà economiche
complessive del settore lattiero-ca-
seario dell’ultimo decennio, deve
affrontare specifiche sfide di tipo
logistico e tecnico.
❚Pasture influence
on fatty acids
profile of mountain
farms bovine milk
SUMMARY
Introduction - Milk and dairy pro-
ducts are considered an important
source of fatty acids in the diet that
varies in quantity according to the
methods of bovine rearing and fee-
ding. In the last decades, evaluation
of the fatty acid composition in bo-
vine milk has been the subject of
several studies for the beneficial ef-
fects on human health. The aim of
this study is to quantify satured and
unsatured fatty acids by indirect
estimation with Infrared spectro-
scopy in bovine bulk tank milk samples collected in typical
mountain farms, in particular to confirm the effect of pastu-
re period on the milk composition.
Materials and methods - The 189 milk samples were collec-
ted from 37 dairy farms representative of the typical “farm”
in a mountainous area of North Italy (Valle Camonica):
small (12 heads/farm on average), prevalence of Brown Alpi-
ne and Red Piebald, low production level (about 16
Lt/head/die), mainly fed with local hay, integration of feed-
stuffs under 30% of the feed ration, without maize silage.
The samples were collected directly from farms (first period,
April-May 2015) or from pasture areas (second period, June-
July 2015).
Results and discussion - Our observations confirm the
change in milk composition related to the grazing period as
well with the increased content in fat and unsatured fatty
acids and, in particular, the “grazing effect” was statistically
confirmed by a significant increase in the unsatured/satured
fatty acids ratio: 0.42 (SD 0.025) in farms vs 0.57 (SD 0.048)
in pasture areas, as already highlighted by other studies ma-
de with reference method.
Conclusions - The Fourier transform infrared spectroscopy
analytical method, easy to use and cheap, although the indi-
Note:
M: valori inerenti il periodo di stabulazione in valle
AP: valori inerenti il periodo di stabulazione in alpeggio
DS: deviazione standard
*Valore espresso come Media Geometrica
G. Bolzoni et al. Large Animal Review 2017; 23: 163-168 167
Grasso (g/100 g) 4,133 (0,475) 4,480 (0,643) Sì
Proteine (g/100 g) 3,471 (0,249) 3,481 (0,201) No
Cellule Somatiche* (cell/ml) 162.850 340.194 Sì
Carica Batterica* (UFC/ml) 117.700 268.900 Sì
Acidi Grassi Saturi (g/100 g) 2,585 (0,344) 2,541 (0,419) No
Acidi Grassi Insaturi (g/100 g) 1,088 (0,166) 1,455 (0,259) Sì
Acidi Grassi Monoinsaturi (g/100 g) 1,004 (0,155) 1,285 (0,246) Sì
Acidi Grassi Polinsaturi (g/100 g) 0,125 (0,019) 0,176 (0,036) Sì
Rapporto Insaturi/Saturi 0,420 (0,019) 0,571 (0,044) Sì
Tabella 2
- Valori medi, deviazione standard e significatività delle differenze tra campioni nel
periodo di stabulazione (M) e nel periodo di pascolo (AP).
Parametro (unità di misura) Media e (DS) Media e (DS) Significatività
M AP Test T (P< 0,05)
Note:
6M: valori inerenti le 6 mandrie che non hanno subito mescolamenti, nel periodo di stabulazione in valle
6AP: valori inerenti le 6 mandrie che non hanno subito mescolamenti, nel periodo di stabulazione in alpeggio
Grasso 4,159 (0,366) 4,589 (0,716) Sì
Acidi Grassi Saturi 2,601 (0,254) 2,600 (0,477) No
Acidi Grassi Insaturi 1,100 (0,163) 1,500 (0,274) Sì
Acidi Grassi Monoinsaturi 1,000 (0,150) 1,353 (0,260) Sì
Acidi Grassi Polinsaturi 0,129 (0,019) 0,182 (0,038) Sì
Rapporto Insaturi/Saturi 0,420 (0,025) 0,573 (0,048) Sì
Tabella 3
- Valori medi, deviazione standard e significatività delle differenze tra campioni nel
periodo di stabulazione (M) e nel periodo di pascolo (AP), limitatamente alle 6 mandrie rimaste
costanti nei due periodi.
Parametro (g/100 g) Media e (DS) Media e (DS) Significatività
Valori 6M Valori 6AP Test T (P< 0,05)
composizione delle mandrie controllate, offrono un’inte-
ressante base informativa per la caratterizzazione del pro-
dotto degli allevamenti nelle aree di montagna ed in parti-
colare per l’apprezzamento dei cambiamenti indotti dal pe-
riodo di pascolo in alta quota. Questa nicchia produttiva ha
una limitata rilevanza dal punto di vista commerciale ri-
spetto al complesso del settore lattiero-caseario del nord
Italia, ma rappresenta un elemento di primaria importanza
per l’economia locale, anche per le sue ricadute in termini
culturali, sociali e di tutela ambientale. In linea generale ri-
teniamo di aver evidenziato che le specifiche, e spesso diffi-
coltose, condizioni gestionali dei piccoli allevamenti mon-
tani consentono di ottenere un prodotto di qualità com-
plessivamente rilevante, adatto alla trasformazione in pro-
dotti caseari tradizionali con alcuni punti di forza ed alcuni
punti critici rispetto al prodotto tipico dell’allevamento in-
tensivo di pianura. Per quanto riguarda la composizione in
acidi grassi, riteniamo di poter concludere che il regime di
pascolo in alta quota costituisce un fattore di significativo
miglioramento del rapporto tra le componenti “desiderate
ed indesiderate” del latte in relazione alla qualità e salubrità
dei prodotti derivati. Questo tipo di osservazioni conferma
quanto già evidenziato da altri Autori con studi in molti ca-
Bolzoni_imp_ok 03/10/17 16:08 Pagina 167
168 Stima dell’effetto pascolo sul profilo in acidi grassi del latte bovino in allevamenti di montagna
rect evaluation of the milk composition, produces results
and information useful to support the quality of milk and
milk products in mountain areas, also for larger projects in
the future.
KEY WORDS
Milk, fatty acids, FTIR, pasture.
Bibliografia
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Bolzoni_imp_ok 03/10/17 16:08 Pagina 168
Bolzoni_imp_ok 03/10/17 16:08 Pagina 169
Bolzoni_imp_ok 03/10/17 16:08 Pagina 170
Garantire il benessere nell’allevamento
dell’asino per la produzione di latte:
un’analisi della legislazione
L. GALLINAa§, F. SAVINIa§, A. LAVAZZAb, P. DI MARCOc, R. PULEIOc, M. ROCCAROa,
G. PURPARIc, A. GUERCIOc, A. SCAGLIARINIa
a Departement of Veterinary Medical Sciences - Alma Mater Studiorum - University of Bologna,
Via Tolara di Sopra, 50 - 40064 Ozzano Emilia (Bologna) Italy
b Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna (I.Z.S.L.E.R.)
Via Bianchi, 7/9 - 25124 Brescia (Italy)
c Istituto Zooprofilattico Sperimentale della Sicilia “A. Mirri” - Via Gino Marinuzzi, 3 - 90129 Palermo (PA)
§ These authors equally contributed to the paper
L. Gallina et al. Large Animal Review 2017; 23: 171-174 171
Bovine papillomatosis in Sicily:
virus identification, diagnosis and co-infections
N
Autore per la corrispondenza:
Laura Gallina (laura.gallina@unibo.it).
SUMMARY
Introduction - Bovine papillomatosis is an infectious disease distributed worldwide among cattle herds consisting of hyper-
proliferative lesions affecting cutaneous tissue and mucosa. The disease, known as papillomas, may occur in different part of
the body. Bovine papillomaviruses (BPVs) are non-enveloped double stranded DNA viruses belonging to the Papillomaviri-
dae family, a group of viruses that infect all amniotes. Despite the benign characters of the lesions, in some cases bovine pa-
pillomatosis may dramatically reduce the productive performances when the lesions are extensive over the body involving
teats and udders.
Aim - This work was particularly aimed at the identification and characterization of BPVs circulating in Sicily in order to ob-
tain autovaccine against BPVs.
Material and methods - Thirty-one epithelial lesions, collected from cattle, clinically diagnosed as papillomas, were referred
to the Laboriatories of Istituto Zooprofilattico Sperimentale of Sicily.
Samples were analyzed by electron microscopy, PCR amplification, Rolling Circle Amplification, sequencing, negative staining
electronic microscopy and standard histological examinations.
Results and discussion - The investigation revealed the presence of different BPV types 1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 12, mostly
in the form of co-infections. Besides BPV amplification, zoonotic viruses belonging to the genus parapoxvirus within the
Poxviridae family were screened and detected in 11 samples. This is the first study performed in Sicily, reporting a wide diver-
sity of BPV genera and types circulating in the territory.
Conclusion - The data collected showed the necessity of proper diagnosis to produce an effective vaccine while virus charac-
terization is important to know the epidemiological scenario of the region and set appropriate control measures and reliable
diagnostic methods for bovine papillomatosis. The presence of co-infecting zoonotic viral agents underlies the need for a bet-
ter understanding of the possible interactions between the two viruses and suggests the importance of identifying proper meas-
ures to prevent the infection to humans.
KEY WORDS
Papillomavirus; diagnosis; autovaccine; Parapoxvirus; co-infections.
INTRODUCTION
Bovine papillomaviruses (BPVs) are non-enveloped ds DNA
viruses causing hyper proliferative benign tumors localized
on skin and mucosa of cattle.
According to Bernard and colleagues (2010), papillomavirus-
es responsible for bovine infections are classified in four gen-
era: Deltapapillomavirus (BPV-1/2/13), Epsilonpapillomavirus
(BPV-5/8), Xipapillomavirus (BPV-3/4/6/9/10/11/12) and
DyoXipapillomavirus (BPV-7) but the development of sensi-
tive detection methods and more intensive research on ani-
mal papillomaviruses have allowed the identification of nov-
el viral types. Further eight putative BPVs have recently been
identified: BPV-14 classified as Deltapapillomavirus, BPV 15,
17, 20 classified as Xipapillomavirus, BPV 16, 18 classified as
Dyokappapapillomavirus and BPV 19 and 21 which have yet
to be assigned to genera and subgroups (Papillomavirus Epis-
teme (PaVE); http://pave.niaid.nih.gov). BPVs infect ker-
atinocytes inducing epithelial papillomas while Deltapapillo-
maviruses determine fibropapillomas of paragenital areas,
skin, alimentary tract and urinary bladder. Cutaneous lesions
normally regress in healthy animals, but they can also result
in economic losses in both beef and dairy production. Papil-
lomas in teats and udders hamper cow milking, and may pro-
mote secondary infections leading to mastitis.
At present no BPV treatment is available and application of
surgical excision of papillomas is not affordable in large herds
with a high prevalence. Despite a number of studies on vac-
cines, there are no commercial formulations against BPVs.
The presence of multiple BPV types and variants in the same
Gallina_imp_ok 03/10/17 16:09 Pagina 171
172 Bovine papillomatosis in Sicily: virus identification, diagnosis and co-infections
lesion have been already reported2,3,4 and suggests the need of
multivalent VLP (virus like particles) vaccines. Otherwise
herd specific vaccine seems to be the easiest practice to confer
protection against the disease caused by different BPV types
even though its efficacy needs to be experimentally proved.
In this light the knowledge of the BPV epidemiological situa-
tion in an area is critically relevant to set up proper control
measures. The aim of this study was to investigate the aetio-
logic agents causing bovine papillomatosis in clinical speci-
mens collected from different provinces in Sicily. Together
with BPVs, the presence of zoonotic parapoxviruses was in-
vestigated as a previous study5reported the co-infection of Pa-
pillomaviridae and Poxviridae in cutaneous lesions of domes-
tic ruminants in Sicily. The parapoxvirus genus comprises two
viral species infecting cattle, Bovine papular stomatitis virus
(BPSV) and Pseudocowpox virus (PCPV). The viruses infect
via damaged skin and give rise to proliferative and/or ulcera-
tive lesions of the skin and mucosae and can infect humans.
MATERIALS AND METHODS
A total of thirty-one clinically diagnosed papillomas were
collected from teats, udders, head and neck by field veteri-
nary practitioners in different provinces of Sicily. Samples
were submitted to Istituto Zooprofilattico Sperimentale Lab-
oratories (IZS) of Sicily for the production of BPV Herd-
Specific Vaccines.
Negative staining Electronic Microscopy (EM) was per-
formed on each sample.
Standard histological examinations were conducted on 18
formalin fixed samples that were processed and stained by
hematoxylin and eosin method (HE). Slides were evaluated
microscopically and histopathologic findings were classified
as fibroapilloma or papilloma.
Viral DNA was purified from frozen pathological samples
(NucleoSpin® Tissue, MN) according to manufacturer’s in-
structions. The quality of the extracted DNA was checked
through the amplification of the bovine β-globin gene. Diag-
nostic PCR and RCA followed by sequencing and enzymatic
digestion were performed to detect and identify the BPV gen-
era and types in each sample. A subset of BPVs type and
genus specific primer pairs were used as already described by
Savini and collaborators6. Amplification was also performed
by multiply-primed Rolling-Circle Amplification (RCA) us-
ing the Illustra TempliPhi 100 amplification kit (GE Health-
care, Little Chalfont, UK) following the protocol previously
described by Rector and collaborators7. A digestion with re-
striction enzymes BamHI and HindIII was subsequently per-
formed to confirm the presence of DNA fragments consistent
with the length of a papillomaviral genome. All specimens
were also subjected to amplification by Pan-parapoxvirus
primers8to detect parapoxviruses (PPV) known to be en-
demic in the area. Purified PCR amplicons were sequenced
using the Sanger method (Perkin-Elmer Applied Biosystems)
and chromatograms were assembled (4 Peaks 1.7.1).
RESULTS
Negative-staining EM showed the presence of particles re-
sembling BPVs in size and shape in thirty samples out of
thirty-one while not any parapoxvirus particle was identified
with this technique. PCR showed a single infection with BPV
in only three lesions while the remaining 28 harbored two (6
lesions) or three (22 lesions) different BPV types. Sequence
analysis and RCA results demonstrated the presence of BPV
1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 12. In particular, twenty-nine sam-
ples harbored BPV1 or 2 (Deltapapillomavirus genus) con-
firming to be the most widespread BPV types in cattle. A
97% identity with the nucleotide sequence of BPV-4 (Gen-
Bank accession n. X05817.1) was shown in one sample
through sequencing of the 400 base pair amplified fragment
of the L1 gene. BPV-5 and BPV-8 belonging to the genus Ep-
silonpapillomavirus were detected in three different samples
while Xipapillomavirus was the second genus mostly detect-
ed in the cutaneous samples (20/31) and BPV-6 was the most
represented (11/20). DyoXipapillomavirus (BPV-7) was de-
tected in seven out of thirty-one specimens. In addition,
RCA amplification and digestion lead to the identification of
patterns consistent in size with BPVs in 27/31 samples. Re-
striction enzyme profiles of digested RCA product allowed
the identification of BPV type in 8 samples while in the oth-
er 19 samples the results were not always in accordance with
PCR findings, as co-infections were detected only in few cas-
es, but most of the time pattern was consistent with the pres-
ence of only one papillomaviral genome.
Coinfection with zoonotic parapoxvirus was detected in 11
samples, sequencing of the PCR products identified 7
PCPV and 4 BPSV. Histologically, lesions were classified as
fibropapillomas (14 samples) and papillomas (4 samples).
Various degrees of hyperplasia of the epidermis with irreg-
ular papillary projections into the dermis were observed in
all samples. Papillomas showed a core of dermal stroma
that supported the proliferating epithelium. Capillaries
within the dermis were dilated and congested. The stratum
corneum exhibited variable degrees of orthokeratotic to
parakeratotic hyperkeratosis. The cells of upper spinous
layer showed swollen lightly basophilic cytoplasm (bal-
looning degeneration), eccentric pyknotic nuclei and a per-
inuclear halo, referred to as koilocytes, the hallmark of BPV
infection. The stratum granulosum had large, variably sized
and shaped basophilic keratohyaline granules, often larger
than normal. Microscopic lesions typical of fibropapillo-
mas showed acanthosis, hyperkeratosis and down growth
of rete ridges, but dermal proliferation was the main fea-
ture (Figure 1). The proliferating cells were large, plump fi-
broblasts, arranged in haphazard whorls and fascicles. The
epidermal proliferation was minimal and only as slight
acanthosis and accentuation of rete pegs was seen.
Despite PCR amplification of zoonotic poxviruses in 11 sam-
ples, histologic examination revealed typical signs of para-
poxvirus infection only in one PCPV and one BPSV-positive
fibropapilloma samples: keratinocyte ballooning degenera-
tion was prominent and scattered, large, eosinophilic cyto-
plasmic inclusions were observed (Figure 2).
DISCUSSION
In this study the identification of the aetiological agents of
clinically diagnosed papillomavirus lesions collected in dif-
ferent cattle farms of Sicily was performed. The presence of
the virus is usually assumed by clinical examination only,
Gallina_imp_ok 03/10/17 16:09 Pagina 172
L. Gallina et al. Large Animal Review 2017; 23: 171-174 173
while studies investigating BPVs’ diversity are sporadic
therefore scarce information about distribution of types in
Italy is available9,6.
In all cases the specimens collected were referred as “papillo-
ma” because the animals were showing multiple proliferative
lesions disseminated throughout the whole body and may
involve a single animal or most of the herd. To date, different
papillomavirus types and genera have been identified as
causative agents of papillomas, in particular 21 different BPV
types belonging to five different papillomavirus genera have
been identified. The combination of different amplification
strategies and sequencing permitted to detect most of the
BPV types and genera officially classified by ICTV. Nu-
cleotide sequencing confirmed the presence of BPV-4 in one
papilloma. This infection was reported by Borzachiello and
collaborators10 and was associated to Oesophageal papillo-
mas in cattle that feed on bracken fern present in the south
of Italy. BPV-4 and BPV-5 weren’t detected in other two re-
cent studies aimed at identification and characterization of
BPV types circulating in North west Italy9and in Emilia-Ro-
magna region6but we don’t know if it is imputable to differ-
ent diagnostic methods used for the investigations or to the
real different widespread of the two BPV types on the Italian
territory. Except for two samples, at least two but more fre-
quently three different type of BPVs were detected in the le-
sions confirming that co-infection of different BPVs in the
same specimen is a frequent finding in field samples. It re-
mains a subject of speculation whether the warts are caused
only by one or more BPVs type, but it is remarkable the need
of a proper diagnostic tool able to detect all different BPVs
especially when no information are available on the viral
types circulating in a certain area. At present mixed BPVs in-
fection have been discovered using generic or genus specific
primers that may fail in the detection of novel BPVs. RCA is
a sensitive method for the detection of novel BPVs and the
use of specific primers is not requested, but after amplifica-
tion digestion by restriction enzyme, cloning and sequencing
are frequently needed for the BPV type identification, be-
coming a labor-intensive and time consuming technique for
routine diagnosis. On the other hand, the identification of
specific viral types present in the papillomatous lesions is
fundamental for appropriate vaccines development like
herd-specific vaccination. In particular, therapeutic vaccine
are requested from the farmers in Sicily because despite the
Figure 1
- Koilocytes (ellipse), basophilic keratohyaline granules in a lesion classified as papilloma (left). Wartlike filiform exophytic prolife-
ration of hyperkeratotic stratified squamous epithelium supported by mature fibrovascular stalks in a fibropapilloma (right). H&E. Bar= 50 µm.
Figure 2
- Ballooning degeneration of keratinocytes of stratum spinosum containing basophilic intracytoplasmic inclusion bodies (ellipse)
typical of parapoxvirus infection. H&E. Bar= 50 µm.
Gallina_imp_ok 03/10/17 16:09 Pagina 173
174 Bovine papillomatosis in Sicily: virus identification, diagnosis and co-infections
proposal of several vaccine models11,12 as no commercial
products are available to prevent BPV infections. As ob-
served by Batista and colleagues13, no connections were
demonstrated among involved viral types and various clini-
cal and histopathological findings particularly in case of
coinfections. Deltapapillomavirus have always been related to
cutaneous fibropapilloma and Xipapillomaviruses to epithe-
lial papilloma14 moreover Grindatto and collaborators9clas-
sified BPV1 infected samples as squamous papilloma and
specimens coinfected by BPV1 and other BPV types as fi-
bropapilloma. The finding of BPV1 or BPV2 in 93.5% of the
specimens confirmed the histological diagnosis of fibropa-
pilloma. Moreover, despite the low sensitivity of ME, nega-
tive staining was able to detect papillomavirus particles in all
samples except one suggesting feasibility of therapeutic vac-
cine preparation even though its efficacy in inducing early
regression of warts is still questionable15. The observation of
eosinophilic inclusion bodies representing the viral factories
of parapoxviruses, suggests the possibility that these zoonot-
ic agents coinfecting papillomatous lesions are biologically
active. Up to now the detection of viruses belonging to the
Poxviridae family was obtained only by molecular biology.
Even though the virus viability still needs to be proved these
data poses major questions about different viruses’ interac-
tion, prevention of zoonotic transmission and supports the
recommendation for clinical diagnoses by laboratory analy-
ses before preparation of herd specific vaccine.
CONCLUSION
This is the first study aiming at detection and characteriza-
tion of BPVs circulating in Sicily. The work confirms what
was previously observed about the high frequency of co-in-
fections in papillomatous lesions of cattle and demonstrates
the wide diffusion of different BPVs on the Italian territory
remarking the need of a standard rapid diagnostic tool for
identification of BPV types in cattle infections. Moreover,
clinical diagnosis needs to be confirmed by laboratory analy-
sis in light of the further reporting of the presence of zoonot-
ic viruses coinfecting papillomatous lesions in cattle and rep-
resenting a potential source of infection for humans when
animals are handled.
CONFLICT OF INTEREST STATEMENT
The research leading to these results has received funding
from the Italian Ministry of Health Ricerca corrente (RC IZS
Si 07-14).
None of the authors of this paper has a financial or personal
relationship with other people or organizations that could
inappropriately influence or bias the content of the paper.
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Gallina_imp_ok 03/10/17 16:09 Pagina 176
Garantire il benessere nell’allevamento
dell’asino per la produzione di latte:
un’analisi della legislazione
M.H. OWEN1, R. DAVIES1, C.E. BEARD1, A.M. OLVERA RAMIREZ1,2, N.R. MCEWAN1
1 IBERS, Aberystwyth University, Aberystwyth, SY23 3DA, Wales
2 Facultad de Ciencias Naturales, Autonomous University of Queretaro, Santiago de Querétaro, Mexico
M.H. Owen et al. Large Animal Review 2017; 23: 177-182 177
Interbreed variation
in craniometrical parameters in sheep
N
Autore per la corrispondenza:
N.R. McEwan (n.mcewan@rgu.ac.uk).
SUMMARY
This work is the first investigation to compare craniometrical parameters between different breeds of sheep. Nine breeds or
cross-breeds of sheep were studied, with all measurements being carried out on live animals at times of routine animal hus-
bandry. Although a number of the variables were not significantly different between breeds, examples of interbreed differences
were found. This was true for both individual measurements of lengths and widths, as well as for indices derived by pair-wise
comparisons of individual widths or lengths. In some cases differences indicated that the cross-bred lambs shared a greater
similarity to one of the parental breeds relative to the other parental breed; e.g. the width between the ears relative to the length
from the nuchal crest to the rostral aspect of the upper lip, where Texel x Wiltshire cross lambs had ratios more similar to the
maternal Wiltshire than they had to the paternal Texel line. In other examples the cross-breeds had values which were inter-
mediate between the two parental lines; e.g. the width at the rostral aspect of the diastema relative to the length from the nuchal
crest to the rostral aspect of the upper lip, where Texel x Wiltshire cross lambs had ratios intermediate between the maternal
Wiltshire and the paternal Texel lines. For one of the ratio values (the length from the nuchal crest to the rostral aspect of the
upper lip, relative to the width at the caudal aspect of the diastema) it was particularly interesting to note that this was a trait
where the male Suffolk x Brecknock Hill Cheviot cross lambs were more similar to the Suffolks (i.e. the paternal line), but the
females cross lambs were more similar to the Brecknock Hill Cheviots (i.e. the maternal line). It is anticipated that this work
will provide implications for using craniometrical parameters as a future husbandry tool.
KEY WORDS
Sheep; craniometrical parameters; interbreed comparisons.
INTRODUCTION
Recording data from domesticated animals is a key compo-
nent of modern husbandry and welfare for both agricultural
animals and pets. Examples of measurements routinely tak-
en vary between species but include variables such as: birth-
weight; weaning weight; withers height, etc.
Craniometry is a long-established discipline in anthropology
as evidenced by a book review by Oetteking from over 100
years ago1. Although originating in human biology it has been
applied to other species. Particularly in dogs, measurements
have been made regarding variables in skull shape (e.g.2),
with skull measurements being used for purposes such as his-
torical breed changes, behavioural traits and general health.
For example, studies have shown historical changes to skull
shapes of St Bernards3as well as links between head measure-
ments and dog trainability4. Based on cephalic indices dog
breeds can be classified into 3 general groups; brachycephalic
(e.g. pugs with relatively flat faces), dolichocephalic (e.g. grey-
hounds with relatively long faces) or mesaticephalic (e.g. col-
lies with intermediate length faces). However, recently other
skull diversity factors have also been studied5.
There has been interest in skull measurements in ruminants,
e.g. Spanish ibex6, goats7-9, roe deer10 and sheep. Interest in
sheep heads involved behavioural analysis11 as well as
anatomical abnormalities. There has also been some work
carried out on head measurements as part of larger whole-
body investigations12, limited investigations of comparisons
between two sheep breeds13-14, and work on multiple meas-
urement points for skulls15-17, on single breeds in each paper.
Geographic features of the British Isles have led to numerous
breeds of domesticated animals, often based on thriving bet-
ter on different grounds (e.g. fast-growing animals on low-
lands, versus hardier animals on uplands). This has led to
over 60 native breeds of sheep within the British Isles18, to-
gether with several breeds imported from mainland Europe
and a range of cross-breeds by crossing purebred lines. Ini-
tially geographical isolation and establishing flock books led
to discrimination between breeds based on morphological
characteristics, although more recently they have been dis-
tinguished by genetic markers (e.g.19). Although breed crite-
ria include morphological characteristics such as body size,
there has generally been little consideration of skull shape, as
it probably has little impact on major selection criteria e.g.
meat production or wool quality.
There are few papers including sheep craniometrical meas-
urements, and with two exceptions using two Turkish sheep
breeds13-14, information on interbreed craniometrical meas-
urements has not underpinned publications. This work in-
vestigates head measurements to analyse differences in head
morphology in different sheep breeds, and compares meas-
urements in cross lambs relative to parental lines.
l
Owen_imp_ok 03/10/17 16:12 Pagina 177
178 Interbreed variation in craniometrical parameters in sheep
MATERIALS AND METHODS
Animals used and measurements made
Three different groups of animals had craniometrical meas-
urements taken. In each case all animals within a group had
measurements determined by a single author at a time of
routine husbandry work; prior to livestock being sold at
market. Measurements (see Table 1) were made using meas-
uring tapes rather than callipers, as they allowed rapid but
accurate measurements with minimal stress to animals
which were not used to being handled.
Group 1
- Craniometrical measurements were made using
3 groups of adult ewes (≥18 months); 30 Southdown, 30
Ryeland and 30 Suftex (Suffolk x Texel crosses). All measure-
ments, 3 lengths (L1 to L3) and 2 widths (W1 and W2) were
recorded by a single author (CEB).
Group 2
- Craniometrical measurements were made using
3 groups of adult ewes (≥ 18 months); 20 Wiltshire Horn
(WH), 20 Texel and 20 WH x Texel crosses. All measure-
ments, 3 lengths (L1 to L3) and 4 widths (W1 to W4) were
recorded by a single author (RD).
Group 3
- Craniometrical measurements were made using
3 groups of lambs aged around 6 months; 50 Brecknock Hill
Cheviot (BHC), 50 Suffolk and 50 BHC x Suffolk crosses - 25
males and 25 females per group. All measurements, 5 lengths
(L1 to L5) and 4 widths (W1 to W4) were recorded by a sin-
gle author (MHO).
Statistical Analysis
Within groups data were checked for normal distribution
before interbreed comparisons using F-test supported stu-
dent T-tests. In addition, a series of indices were derived us-
ing one measurement divided by another, and expressed as
the first variable as a percentage of the second. This was per-
formed for all length: width and width: width combinations,
and the statistical analysis repeated as above.
RESULTS
Group 1
Data for adult Southdown, Ryeland and Suftex ewes are
shown in Table 2. This shows that sizes of some features did
not vary significantly between breeds. For example, the
length from the nuchal crest to the rostral aspect of the up-
per lip [L1] was not significantly different between breeds.
However, other measurements did vary, e.g. width between
the ears [W1] was significantly different between breeds.
Furthermore, the length from the caudoventral aspect of the
Figure 1 - Positions at which measurements were made on the
sheep heads. Note that all measurements were made in live animals
but this image uses a skull to make viewing easier.
W1 Width between the ears ab
W2 Width at rostral aspect of diastema - between outer incisors (I4) of left and right jaws c d
W3 Width between caudal aspects of left and right jaws e f
W4 Width at caudal aspect of diastema - between first premolars (P1) of left and right jaws g h
L1 Length from nuchal crest to rostral aspect of upper lip i j
L2 Length from caudoventral aspect of angle of mandible to rostral aspect of lower lip k l
L3 Length from caudal aspect of hinge of lower jaw to nasal tip m n
L4 Length from akrokranion to rostral aspect of upper lip o j
L5 Poll to the rostral aspect of upper lip pj
Table 1
- Variables measured within the current work. The symbols denote the position shown on the image in Figure 1.
Symbol 1 Symbol 2
Owen_imp_ok 03/10/17 16:12 Pagina 178
M.H. Owen et al. Large Animal Review 2017; 23: 177-182 179
mandible angle to the rostral aspect of the lower lip [L2] was
significantly longer in Suftexs (x
–=158 mm, SEM=2.3) than
Ryelands (x
–=140 mm, SEM=3.9), but neither differed signif-
icantly from that of the intermediate Southdowns (x
–=149
mm, SEM=4.1).
Group 2
The data for adult Texel, WH and Texel x WH cross ewes are
shown in Table 3. Differences in absolute values for these
breeds were not significant (P>0.05).
However indices derived from these measurements showed
significant differences. In some indices the parental breeds
were significantly different, but cross ewes had intermediate
values and were not significantly different from either par-
ent. One such an index was the ratio between L1 (length
from nuchal crest to aspect of upper lip) relative to W1
(width between ears).
In other indices, cross ewes were not significantly different
from WHs, but both cross ewes and the WHs were signifi-
cantly different from Texels e.g. L1 (length from nuchal crest
to aspect of upper lip) relative to W3 (width between cau-
doventral aspects jaws).
There were no indices where crosses showed similarity to
Texels but not to the WHs.
One index, W4 (width between caudal aspect of diastema)
relative to L1 (length from nuchal crest to rostral aspect of
upper lip) was significantly different in all 3 breeds, with
values for crossbreds intermediate between those for pure
breeds.
Finally, there was an example where Texels and WHs were
not significantly different, but cross ewes were significantly
different from either pure breed. This was seen when com-
paring W2 (width at rostral aspect of diastema) relative to
W3 (width between caudoventral aspects of jaws). It was as-
sumed that this observation was due to proportions of the
Texel and WH head shapes being similar, but that propor-
tions in crosses were different.
Group 3
All animals were around 6 months old when measured. At
this age some measurements were already different, although
this could be due to developmental differences between
breeds, rather than interbreed differences in mature animals.
For example the width between the ears [W1] differed based
on a combination of breed and gender factors, with male
Suffolks having the largest means (x
–=154 mm, SEM=2.8)
and female BHCs having the smallest mean value (x
–=90
mm, SEM=1.3).
Examples were seen where cross lambs had values more sim-
ilar to those of the BHCs than those of Suffolks. For example
the length from the caudal aspect of the hinge of the lower
jaw to the nasal tip [L3], where differences were not signifi-
cant between crosses and BHCs (P>0.05), irrespective of
gender (mean lengths ranged from 178-181 mm). However
this value was significantly longer in Suffolk females (x
–=187
mm, SEM=1.6), and males (x
–=206 mm, SEM=3.2).
The opposite was seen in other examples. The index derived
by comparing L2 (length from caudoventral aspect of angle
of mandible to rostral aspect of lower lip) relative to W2
(width at rostral aspect of diastema) was not significantly
different (P>0.05) in Suffolk lambs and cross lambs with
males (x
–=26 in both breeds) and females (x
–=25 in both
W1 154 a(2.4) 143 b(1.6) 126 c(2.2)
W2 76 a(1.5) 84 b(2.5) 121 c(2.3)
L1 244 a(3.5) 239 a(5.8) 239 a(2.6)
L2 140 a(3.9) 149 ab (4.1) 158 b(2.3)
L3 175 a(3.8) 173 a(3.4) 208 b(2.3)
W1 - L1 63 a(1.4) 61 a(1.4) 53 b(0.9)
W1 - L2 114 a(3.4) 99 b(2.5) 80 c(1.8)
W1 - L3 90 a(2.4) 84 a(1.8) 61 b(1.1)
W2 - L1 31 a(0.7) 35 b(0.7) 51 c(0.9)
W2 - L2 56 a(1.9) 57 a(1.3) 77 b(1.8)
W2 - L3 44 a(1.3) 49 a(1.1) 58 b(1.2)
W1 - W2 206 a(4.5) 176 b(4.6) 105 c(2.3)
Table 2
- Five measurements (in mm) recorded for the Ryeland,
Southdown and Suftex (Suffolk x Texel) adult ewes (n=30 for each
breed), together with the corresponding indices which have been de-
rived by expressing the size of the first variable as a percentage of the
second variable. SEM values are shown in parenthesis. Values within
rows which are not significantly different (P>0.05) share a superscript.
Ryeland Southdown Suftex
W1 146 a(1.4) 151 a(2.8) 144 a(1.8)
W2 74 a(1.6) 73 a(1.2) 73 a(1.1)
W3 107 a(2.3) 107 a(2.2) 104 a(2.2)
W4 94 a(1.7) 94 a(1.7) 99 a(1.3)
L1 230 a(2.0) 259 a(1.4) 247 a(1.8)
L2 185 a(2.5) 172 a(2.4) 172 a(2.6)
L3 178 a(2.9) 178 a(4.7) 166 a(1.8)
W1 - L1 63 a(0.6) 59 b(1.1) 58 b(0.7)
W1 - L2 79 a(0.9) 88 b(1.7) 84 ab (1.6)
W1 - L3 83 a(1.3) 87 a(2.3) 87 a(1.5)
W2 - L1 32 a(0.6) 28 b(0.5) 30 ab (0.5)
W2 - L2 40 a(0.8) 43 a(0.9) 43 a(1.0)
W2 - L3 42 a(1.4) 42 a(1.3) 44 a(0.7)
W3 - L1 51 a(1.0) 41 b(0.8) 42 b(1.0)
W3 - L2 63 a(1.2) 62 a(1.5) 61 a(1.7)
W3 - L3 66 a(1.5) 61 a(1.7) 63 a(1.1)
W4 - L1 46 a(0.8) 37 b(0.7) 40 c(0.6)
W4 - L2 57 a(1.2) 55 a(1.2) 58 a(1.4)
W4 - L3 60 a(1.6) 55 a(1.8) 59 a(0.8)
W1 - W2 201 a(4.2) 213 a(6.8) 199 a(4.3)
W1 - W3 126 a(2.2) 145 b(3.9) 140 b(3.7)
W1 - W4 140 a(2.9) 165 b(5.2) 148 ab (3.2)
W2 - W3 63 a(1.2) 70 ab (1.8) 71 b(1.2)
W2 - W4 70 a(1.1) 78 b(1.0) 74 ab (0.8)
W3 - W4 112 a(2.0) 115 a(3.0) 106 a(1.8)
Table 3
- Seven measurements (in mm) recorded for the Texel, Wilt-
shire Horn and Texel x Wiltshire Horn adult ewes (n=30 for each
breed), together with the corresponding indices which have been de-
rived by expressing the size of the first variable as a percentage of the
second variable. SEM values are shown in parenthesis. Values within
rows which are not significantly different (P>0.05) share a superscript.
Texel Wiltshire Texel x Wiltshire
Owen_imp_ok 03/10/17 16:12 Pagina 179
180 Interbreed variation in craniometrical parameters in sheep
breeds). However the mean BHC values were significantly
shorter (23 and 21 for males and females respectively).
As well as interbreed differences, gender differences were ob-
served. For example the length from the nuchal crest to the
rostral aspect of the upper lip [L1]. In this case male BHCs
(x
–=224 mm, SEM=2.1) and male crosses (x
–=218 mm,
SEM=2.5) were not significantly different (P>0.05), and fe-
male BHCs (x
–=208 mm, SEM=2.8) and female crosses
(x
–=208 mm, SEM=2.3) were not significantly different
(P>0.05). However both lengths in males were significantly
(P<0.05) longer than in females.
Indices were also observed with differences between breeds
and genders. For example the ratio between W4 (width at
caudal aspect of diastema) and L1 (length from nuchal crest
to rostral aspect of upper lip). Male and female Suffolks,
(x
–=22, SEM=0.7 and x
–=22, SEM=0.7 respectively) did not
differ significantly (P<0.05). Likewise male and female
BHCs, (x
–=20, SEM=0.4 and x
–=20, SEM=0.5 respectively)
W1 154 a(2.8) 131 b(2.1) 102 c(1.8) 90 d(1.3) 138 e(2.2) 112 f(2.6)
W2 44 a(1.12) 39 b(1.0) 36 c(0.7) 32 d(0.6) 41 b(0.7) 38 c(0.5)
W3 97 a(2.0) 87 b(1.7) 78 c(1.3) 74 d(1.2) 84 b(1.6) 69 e(1.2)
W4 58 a(1.2) 55 b(1.4) 44 c(0.9) 41 d(0.8) 49 e(1.1) 42 cd (0.7)
L1 266 a(5.8) 249 b(3.8) 224 c(2.1) 208 d(2.8) 218 c(2.5) 208 d(2.3)
L2 176 a(3.6) 160 bc (2.4) 160 bc (1.8) 156 bde (1.7) 160 cd (2.1) 151 e(1.8)
L3 206 a(3.2) 187 b(1.6) 178 c(1.9) 178 c(1.5) 181 c(2.4) 178 c(1.3)
L4 346 a(4.8) 323 b(3.2) 273 c(3.4) 259 d(3.0) 271 c(4.1) 273 c(2.6)
L5 228 a(2.82) 213 b(2.3) 191 c(1.6) 180 d(1.3) 190 c(1.6) 187 c(1.6)
W1 - L1 58a(1.5) 53 b(1.2) 46 c(0.9) 44 c(0.9) 63 d(1.3) 54 b(1.4)
W1 - L2 88 a(2.9) 83 a(1.9) 64 b(1.3) 58 c(1.0) 86 a(1.7) 74 d(1.9)
W1 - L3 75 a(1.7) 70 b(1.4) 57 c(1.1) 51 d(0.9) 77 a(1.6) 63 e(1.4)
W1 - L4 45 a(0.8) 41 b(0.7) 37 c(0.6) 35 d(0.7) 51 e(1.) 41 b(1.0)
W1 - L5 68 a(1.1) 62 b(1.2) 54 c(1.2) 50 d(0.8) 73 e(1.3) 60 b(1.4)
W2 - L1 17 ab (0.6) 16 a(0.5) 16 a(0.4) 16 a(0.4) 19 c(0.4) 18 bc (0.3)
W2 - L2 26 a(1.0) 25 a(0.6) 23 b(0.5) 21 c(0.4) 26 a(0.5) 25 a(0.4)
W2 - L3 22 ab (0.6) 21 a(0.6) 20 a(0.4) 18 c(0.3) 23 b(0.4) 21 a(0.3)
W2 - L4 13 a(0.3) 12 a(0.4) 13 a(0.3) 13 ab (0.3) 15 c(0.3) 14 b(0.2)
W2 - L5 19 ab (0.5) 19 ac (0.6) 19 ac (0.4) 18 c(0.3) 22 d(0.4) 20 b(0.3)
W3 - L1 37 ab (1.2) 35 ac (0.9) 35 ad (0.7) 36 a(0.7) 39 b(0.8) 33 cd (0.7)
W3 - L2 55 a(1.2) 55 a(1.1) 49 b(1.0) 48 bc (0.8) 53 a(1.1) 46 c(1.0)
W3 - L3 47 a(1.0) 47 a(1.0) 44 b(0.7) 42 c(0.7) 47 ab (1.1) 39 d(0.7)
W3 - L4 28 a(0.7) 27 a(0.6) 29 a(0.7) 29 a(0.5) 31 b(0.7) 25 c(0.5)
W3 - L5 43 ab (0.9) 41 a(1.1) 41 a(0.7) 41 a(0.7) 44 b(0.9) 37 c(0.6)
W4 - L1 22 a(0.7) 22 a(0.7) 20 b(0.4) 20 b(0.5) 22 a(0.5) 20 a(0.5)
W4 - L2 33 a(0.9) 35 a(1.0) 28 bc (0.6) 26 b(0.5) 30 d(0.7) 28 c(0.5)
W4 - L3 29 ab (0.7) 30 a(0.9) 25 c(0.5) 23 d(0.4) 27 b(0.8) 24 cd (0.5)
W4 - L4 17 abc (0.5) 17 ac (0.5) 16 abd (0.2) 16 be (0.4) 18 c(0.5) 16 de (0.3)
W4 - L5 26 a(0.6) 26 a(0.8) 23 b(0.5) 23 b(0.4) 26 a(0.7) 23 (0.4) b
W1 - W2 266 a(6.9) 241 b(5.4) 232 bc (4.3) 223 c(5.8) 286 a(7.6) 266 a(7.3)
W1 - W3 352 a(8.6) 337 a(8.5) 284 b(5.4) 281 b(5.3) 337 a(6.4) 299 b(8.7)
W1 - W4 161 ab (4.7) 152 a(3.5) 131 c(3.3) 122 d(2.3) 165 b(3.8) 162 ab (4.3)
W2 - W3 47 ab (1.2) 46 a (0.8) 47 ab (0.9) 44 a(0.6) 50 b(0.9) 54 c(0.8)
W2 - W4 77 ab (1.4) 71 a (1.3) 81 cd (1.3) 80 bc (1.2) 87 de (1.3) 89 e(1.3)
W3 - W4 167 ab (3.7) 160 a(4.4) 179 c(4.7) 182 c(3.9) 174 bc (3.7) 165 ab (3.4)
Table 4
- Nine measurements (in mm) recorded for the Suffolk, Brecknock Hill Cheviot and Suffolk x Brecknock Hill Cheviot lambs (25 ma-
les and 25 females for each breed), together with the corresponding indices which have been derived by expressing the size of the first va-
riable as a percentage of the second variable. SEM values are shown in parenthesis. Values within rows which are not significantly different
(P>0.05) share a superscript.
Suffolk Brecknock Hill Cheviot Suffolk x Cheviot
Male Female Male Female Male Female
Owen_imp_ok 03/10/17 16:12 Pagina 180
M.H. Owen et al. Large Animal Review 2017; 23: 177-182 181
were not significantly (P<0.05) different. However male
crosses (x
–=22, SEM=0.5) were more similar to Suffolks and
female crosses (x
–=20, SEM=0.5) were more similar to BHCs.
A similar pattern was seen for the index derived from W4
(width at caudal aspect of diastema) relative to L5 (length
from poll to top lip); male crosses being more similar to Suf-
folks and female crosses being more similar to BHCs.
DISCUSSION
Previous studies of sheep craniometrical measurements tend
to have concentrated on using skulls without soft tissue. The
two exceptions to this13-14, were part of a wider range of
measurements across the body. Therefore, as far as we are
aware, this is the first comparison of craniometrical meas-
urements between breeds in a single paper, and the first
analysis of multiple head measurements in live sheep. The
current work uses data collected by three individuals from
different sites. Therefore no comparison has been made be-
tween groups, to avoid potential inter-experimenter varia-
tion. In addition, groups 1 and 2 involved adult animals, but
group 3 used younger animals, again meaning comparisons
across all groups were not appropriate. Repeating measure-
ments by a single experimenter could have allowed compar-
isons between groups, but animals were measured during
routine handling after which some animals were sold, mean-
ing they were not available for retrospective measurements.
The work involved measurements with a tape, rather than
callipers as these sheep were not used to being handled, and
so minimised stress levels with rapid recording of data.
However SEM values within groups (Tables 2 to 4) were all
relatively low, suggesting low variability within single sam-
ples, supporting the assumption that tapes gave adequately
reliability.
The first set of analysis were performed primarily to demon-
strate that differences (e.g. W1) between breeds could be de-
tected using a measuring tape with low variability. Therefore
measurements undertaken were the five which were felt
could be made rapidly and would involve minimal stress to
the animals. Using these values it was possible to demon-
strate that statistically significant (P<0.05) interbreed differ-
ences could be detected in terms of absolute values, and also
in indices derived from these measurements. Moreover, sta-
tistical analysis of these identified statistically significant dif-
ferences. In this example, two of the breeds were unrelated
purebred lines, and the third breed was a cross which was not
derived from either of these. Thus no relationship compar-
isons were performed between the three sets of animals.
The second analysis used two pure breeds; Texels and WHs,
and a cross derived by mating Texel rams and WH ewes. A
further two measurements were included, again ones which
it was felt could be made rapidly and with minimal stress. As
above only adult ewes were used. This group was used to ex-
amine if particular distances or indices in the crosses were
more similar to those seen in specific parents. Two indices
were not significantly (P>0.05) different in the WHs and
crosses, but significantly different (P<0.05) in Texels relative
to the other breeds; ratio of the width between the ears [W1]
relative to width between the caudoventral aspects of jaws
[W3], and ratio of the width at the rostral aspect of the di-
astema [W2] relative to width at the caudal aspect of the di-
astema [W4]. The values in the cross ewes were more similar
to those in WH ewes, suggesting these traits may show a ge-
netic bias.
In this group there were also two examples where crosses had
intermediate values, which were not significantly different
(P>0.05) from either parental breed, but both parental
breeds differed significantly (P<0.05) from each other; ratio
of the length from the nuchal crest to the rostral aspect of the
upper lip [L1] relative to width at the rostral aspect of the di-
astema [W2], and ratio of width between the ears [W1] rel-
ative to width at the caudal aspect of the diastema [W4].
Both of the first two groups used data from adult ewes,
meaning potential gender differences were not considered.
Primarily this was due to individual farms generally have
enough adult ewes available to allow comparisons, but insuf-
ficient adult rams to permit this. In mammals males tend to
be larger and heavier, a factor which prompted the third
study involving males and female from two pure breeds
(BHCs and Suffolks), and crosses produced from BHC ewes
and Suffolk rams. However, routinely insufficient males are
retained for breeding, meaning analysis were restricted to
animals at 6 months, rather than adults as above. A further
two measurements were included at this point, again ones
which could be carried out rapidly and with minimal stress.
The length from the nuchal crest to the rostral aspect of the
upper lip [L1] in male BHC lambs (x
–=224 mm, SEM=2.1)
and male crosses (x
–=218 mm, SEM=2.5) were not signifi-
cantly different, and those in female BHC lambs (x
–=208
mm, SEM=2.8) and female cross lambs (x
–=208 mm,
SEM=2.3) were also not significantly different, suggesting
this length in the cross lambs had more in common with the
equivalent length in BHC lambs of the same sex.
The length from the caudal aspect of the hinge of the lower
jaw to the nasal tip was not significantly different between
the BHC and cross lambs; irrespective of gender. Neither of
these factors was observed in Group 2 study using Texels,
WHs and their cross.
It is worth noting that the mean length from the akrokran-
ion to the rostral aspect [L4] of the upper lip and the mean
length from the poll to the top lip [L5], measurements not
recorded in the previous two groups, both showed signifi-
cant similarity in length in cross lambs of both sexes relative
to the equivalent distance in male BHC lambs.
Some indices provided interesting comparisons for prelimi-
nary comparisons between groups - albeit they were carried
out by different individuals. For example the ratio of width
between the ears [W1] relative to width between the cau-
doventral aspects of left and right jaws [W3], where Texel x
WH crosses were more similar to WHs (maternal line) than
Texels, but BHC x Suffolk crosses (both genders) showed
more similarity to male Suffolks (i.e. the paternal lineage).
The ratio between the length from the nuchal crest to the
rostral aspect of the upper lip [L1] and width between the
ears [W1] was not significantly different (P>0.05) between
Suffolk females and female BHC x Suffolk crosses. This pat-
tern was repeated for the length from the poll to the top lip
[L5] relative to width between the ears [W1], with similar
values between Suffolk females and female BHC x Suffolk
crosses, again similarity with the maternal lineage.
In other cases there was similarity between Suffolk males
and male BHC x Suffolk crosses. There was no significant
difference in ratios between the length from the caudal as-
Owen_imp_ok 03/10/17 16:12 Pagina 181
182 Interbreed variation in craniometrical parameters in sheep
pect of the hinge of the lower jaw to the nasal tip [L3] rel-
ative to width between the ears [W1], and the length from
the caudoventral aspect of the angle of the mandible to the
rostral aspect of the lower lip [L2] relative to width between
the ears [W1].
The ratio of width at the rostral aspect of the diastema to the
length from the caudoventral aspect of the angle of the
mandible to the rostral aspect of the lower lip was similar in
both the Suffolk lambs and BHC x Suffolk crosses. However
the values for the BHCs showed significant (P<0.05) gender
differences. This group identified potential gender differ-
ences for one index; ratio of the length from the nuchal crest
to the rostral aspect of the upper lip [L1] relative to width at
the caudal aspect of the diastema [W4], where male crosses
were more similar to Suffolks (both genders), but the female
crosses more similar to BHCs (both genders).
The full implications of this work are unclear as this is the
first investigation in sheep on this scale. The role of cranio-
metrical parameters in understanding problems in dogs has
been clear for some time (e.g.2) with more detailed recent
studies (e.g.5), particularly the role of head shape to health
issues such as birthing and respiratory problems in brachy-
cephalic dogs. However factors affecting heads are being
implicated as important in understanding pain and stress
in sheep20.
It is in these areas that analysing craniometrical parameters,
and estimating what constitutes normal, may have a future
impact on sheep breeding. In some sheep breeds head size
and shape can pose lambing problems. It has been estimated
that dystocia accounts for around half of deaths in lambs in
the first 72 hours after birth21, i.e. around 7% of lambs born.
Although lambing difficulties vary between breed and are
not restricted to the lamb’s head, these pose major problems
at lambing time, particularly when numbers are considered
at both national and international levels.
Furthermore, some sheep breeds, as with some brachy-
cephalic dogs, are prone to respiratory and breathing prob-
lems e.g. laryngeal chondritis. Therefore an exploration of
a potential linkage between head morphology and traits
such as respiratory problems later in life is worth further
exploration.
In conclusion, this work is the first to carry out interbreed
comparisons of a range of craniometrical parameters in live
sheep. Examples of differences were found between breeds,
including some suggesting that some parameters in the cross
breeds are more similar to one of the parental breeds than
the other, and also examples of intermediate mean values.
Moreover, differences within breeds could be seen between
male and female lambs, including an example where the
male cross lambs showed more similarity to the paternal
breed and the ewe lambs showed more similarity to the ma-
ternal breed.
Nine breeds, or crosses of breeds were studied in the current
work, and up to nine measurements were made for each
breed. These data for variation between breeds in terms of
head measurements and craniometrical indices derived from
them, may argue for head measurements becoming a topic
worth investigating in more detail, using more measure-
ments than those used in the current work. Moreover the
range of domesticated breeds which exist internationally,
coupled with other species in the Ovis genus and other gen-
era in the Caprinae Subfamily, argue that there is scope for
further craniometric measurements in other animals to un-
derstand these relationships.
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2. Onar V (1999) A morphometric study on the skull of the German shep-
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4. Helton WS (2009) Cephalic index and perceived dog trainability. Behav
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Owen_imp_ok 03/10/17 16:12 Pagina 183
Owen_imp_ok 03/10/17 16:12 Pagina 184
Monitoring of selected milk quality
parameters during lactation of
a Mediterranean dairy sheep breed
T.G. BONTINIS*, E.C. PAPPA*, K. SOTIRAKOGLOU**, A.C. PAPPAS***,
E. TSIPLAKOU***, G. ZERVAS***
*Dairy Research Department, Directorate General of Agricultural Research, Hellenic Agricultural Organization
DEMETER, Katsikas, 45221 Ioannina, Greece
**Department of Plant Breeding and Biometry, Faculty of Crop Science, Agricultural University of Athens,
Iera Odos 75, 11855 Athens, Greece
***Department of Nutritional Physiology and Feeding, Faculty of Animal Science and Aquaculture,
Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece
T.G. Bontinis et al. Large Animal Review 2017; 23: 185-188 185
Corresponding author:
Eleni C. Pappa (pappa.eleni@yahoo.gr)
SUMMARY
Introduction - Sheep and goats are the most efficient transformers of low quality forage into high quality animal products
(milk and meat) with distinguished characteristics. Sheep and goat milk production in several Mediterranean countries is
mainly taking place in less favoured, rural and mountainous areas. In Greece, sheep production is of high importance for
mountainous, arid, semi-mountainous and semi-arid areas, in which sheep are exploited under extensive or semi-extensive
management regimes. In those areas, the use of autochthonous breeds is preferred. Arta sheep breed (known as Frizarta) is a
local breed reared in the North-Western areas of Greece. Milk composition is a key factor that influences milk quality, and in
turn the ratio processed dairy products and farm economics.
Material and methods - The aim of the present study was to monitor selected milk quality parameters during lactation of the
Arta sheep breed in a less favoured, rural and mountainous area of North-Western Greece. A total of 840 randomly selected
ewes from the Arta breed (8 different flocks), managed under semi-extensive production system, were examined during sever-
al lactation stages (months) over three consecutive years. Milk was analysed for somatic cell count and fat, protein, lactose,
solids-non-fat content.
Results and discussion - Lactation stage significantly altered milk composition but not somatic cell count. Lactose was nega-
tively correlated with somatic cell count as well as fat and protein content and positively with solids-non-fat content. There was
a negative correlation between somatic cell count and solids-non-fat content. The average annual milk yield for the first, sec-
ond and third year was 250, 260 and 280 kg respectively with no statistical significant differences among years.
Conclusions - The results indicate that milk quality of the Arta breed is good. The good milk quality can support the produc-
tion of local traditional cheeses. Locally made dairy products of added value can in turn positively affect the income of farm-
ers and contribute to the endurance of rural human population in less favoured areas.
KEY WORDS
Arta breed, dairy ewes, less developed, Mediterranean, milk composition.
INTRODUCTION
At the European level, the dairy sheep population is con-
fined, at a large part, to the Mediterranean countries. The in-
tensification and dairy specialization processes have taken
place throughout the whole Mediterranean area especially in
countries such as France and Spain and to a lesser extent in
Greece1. Sheep and goat milk production in several Mediter-
ranean countries is mainly situated in less favoured, rural
and mountainous areas1. More specifically, Greece, and Italy
and a few other countries, are characterised as countries with
a high proportion of mountains considering the high per-
centage of utilised agricultural area designated as less
favoured areas2.
Ewe’s milk is widely used in the Mediterranean and Balkan
regions for the production of cheese, yoghurt and other
dairy products. Furthermore, milk composition is of great
importance since it affects the quality and determines the ra-
tio “processed product/milk” and therefore the cost. Improv-
ing milk yield and quality of sheep milk in less developed,
rural and mountainous areas is of paramount importance in
order to increase farming economics. The composition of
sheep milk varies widely in relation to breed, nutritional and
environmental factors, stage of lactation, parity and sea-
son3,4,5,6,7,8,9. More specifically, in the semi-extensive produc-
tion systems of dairy sheep in Greece, the feeding is based
mainly on grazing native pastures whose composition de-
pends on climatic conditions and seasonality. In such feeding
regimes, the animal nutritional requirements may be under
or over met which affect the milk yield and chemical com-
position10. Moreover, in these farming productions systems
the milking is, in most cases, operated manually which also
l
4 BONTINIS_imp_ok 03/10/17 16:17 Pagina 185
Statistical analysis
Statistical analysis was performed using statistical package
SPSS (version 14.0, Chicago, IL, USA) and results are pre-
sented as mean ± root mean square error. Somatic cell
counts were log10 transformed. Kolmogorov-Smirnov test
revealed that all data followed normal distribution. Differ-
ences in chemical composition and count of somatic cells of
ewe’s milk between lactation stages over the three consecu-
tive years were tested using repeated measures analysis of
variance (ANOVA) with two factors, lactation stage and year.
Multiple comparisons were obtained using Tukey’s test.
Pearson’s correlation coefficients were used to determine re-
lationships between the examined physicochemical proper-
ties. For all tests a P-value of less than 0.05 was considered
significant.
RESULTS AND DISCUSSION
Milk characteristics
Mean values of milk composition and SCC at different lacta-
tion stages (months) are shown in Table 1. Lactation had a
significant effect on all main constituents (P<0.05), except
lactose and LogSCC. In detail, the fat content of milk exhib-
ited a gradual decrease from the beginning of the lactation
period, reaching a minimum in the middle of lactation
(spring) followed by a constant increase until the end of lac-
tation in July. A similar variation pattern for fat composition
has an impact on milk quality. Thus, the aim of the present
study was to monitor the milk quality of the Arta sheep
breed which is farmed in a less favoured, rural and moun-
tainous area of North-Western Greece.
MATERIALS AND METHODS
Characteristics of Arta breed
The Arta sheep breed, also known as Frisarta, was developed
in Greece more than 40 years ago based on a breeding pro-
gram supported and monitored by the Ministry of Rural De-
velopment and Food aiming to the genetic improvement of
the indigenous sheep population. It was developed with the
objective of improving milking potential and prolificacy of
indigenous sheep. Following an intensive breeding selection
program that involved crossing East Friesian rams with an
indigenous sheep population.
The greatest concentration of these sheep is on the plains of
Arta in Western Greece. Arta ewes are of large size and body
weight (63.3±7.8 kg) with white hair and a long thin tail11,12.
It is a dairy breed with average annual milk yield of about
230-250 kg/head, making it one of the highest milk produc-
ing breeds in Greece13. Milk yield is peaked in April and re-
duced by late July to a level which is no longer commercially
viable. The average herd size ranges between 50 to150 ewes
and is considered relatively small. Husbandry and herd man-
agement (feeding regiments, health status and confine-
ments) are evaluated as satisfactory.
Management of the sheep
The study was conducted on the plains of Arta
prefecture, located in North-Western Greece. A
total of 840 dairy sheep of Arta breed, from 8 dif-
ferent flocks of the same typical feeding and man-
agement practices, were used. Lambing started in
mid-November and lasted about 2 weeks. The
lambs were then suckling their mothers approxi-
mately 45 days. The milking of ewes started at the
beginning of January.
Sheep nutrition was based mainly on supplemen-
tary feeding during the winter months, up to mid-
March while from April onwards grazing native
pastures was the only source of feed. The native
pastures consisted predominantly of graminaceae
and leguminosae and to a smaller extend of papil-
ionacea and compositae. The supplementary
feeding consisted of concentrates plus alfalfa hay
and straw. The composition of the concentrate di-
et (g/kg) which was given to sheep was the follow-
ing: maize, 450; barley, 154; wheat middlings, 235;
soybean meal, 130; CaCo3, 24; NaCl, 5 and vita-
min and mineral premix, 2.
Milk records and samples were obtained fort-
nightly for three consecutive lactation periods.
Samples of milk were analysed for fat,protein, lac-
tose, solids-non-fat (SNF) using i.r. spectropho-
tometer (Milko Scan, model 6000; Foss Electric,
Hillerod, Denmark). Furthermore, somatic cell
count (SCC) in milk was performed by a Fosso-
matic 6000 (Foss Electric) apparatus using the
standard of the International Dairy Federation14.
186 Monitoring of selected milk quality parameters during lactation of a Mediterranean dairy sheep breed
Data are means of eight replicates (n=8). Means within a column, within a main effect
comparison, with different superscripts are different at P<0.05. NS: non significant; SNF:
solids-non-fat; SCC: Somatic cell count; RMSE: root mean square error.
Lactation
(month)
January 6.75ab 5.84b4.85 11.37b5.92
February 6.90b5.81b4.92 11.39b5.94
March 6.19a5.68ab 5.03 11.35b5.89
April 6.20a5.46ab 5.02 11.19ab 5.90
May 6.18a5.53ab 4.94 11.16ab 5.95
June 6.79ab 5.39a4.88 11.04a6.07
July 7.12b5.56ab 4.83 11.10ab 6.00
Year
1st 6.65 5.51a4.92 11.14a6.07
2nd 6.59 5.66b4.93 11.21ab 5.92
3rd 6.59 5.73b4.93 11.33b5.86
RMSE 0.33 0.35 0.14 0.34 0.16
Source of
Variation P value
Lactation 0.036 0.008 NS 0.001 NS
Year NS 0.002 NS 0.008 NS
Lactation 0.037 NS NS 0.032 0.029
x Year
Table 1
- Composition and count of somatic cells of ewe’s milk per lactation
over a period of three consecutive years.
Factor Fat Protein Lactose SNF LogSCC
studied (%) (%) (%) (%)
4 BONTINIS_imp_ok 03/10/17 16:17 Pagina 186
Protein 0.33***
Lactose -0.45*** -0.22**
SNF 0.16* 0.82*** 0.19*
LogSCC 0.27*** -0.01 -0.32*** -0.16*
T.G. Bontinis et al. Large Animal Review 2017; 23: 185-188 187
has been reported by Voutsinas et al.15. Average fat content
was lower compared to the one reported by Boyazoglu and
Morand-Fehr7for the Greek breeds of Chios, Vlahiko,
Karagouniko and Attiki, but higher than that reported for
milk of some breeds in Mediterranean countries16,17,18. Simi-
lar results have been reported for the Arta breed and Chios
breed, respectively11,19. Lower fat content in spring was due to
lower NDF, ADF content of the diet (grass).
Protein content of milk progressively decreased to a mini-
mum in June followed by a gradual increase until the end of
the lactation. Season can affect milk fat and protein and in
hot, humid months fat and protein content can be depressed
possibly attributed to changes of feed intake patterns which
may be lower in summer due to changes in weather and tem-
perature. The above results are in agreement with the results
reported by Papavasiliou 11 for the same breed, similar to the
ones reported for the Greek breed of Chios19 and for Mass-
ese ewes16, but lower compared to those reported for other
Greek breeds15,20. Overall, average protein content was high-
er than that reported in foreign breeds16,21.
Lactose content of milk, did not exhibit statistically signifi-
cant alterations as lactation progressed although was 3%
higher in spring compared to summer values. A similar pat-
tern has been reported in similar studies22,23. Similar results
were reported for the Chios breed by Ploumi et al.19. Average
lactose content was similar to those reported for various
Greek11,15,20 and foreign breeds16.
The SNF content progressively decreased to a minimum in
June compared to the beginning of lactation. The results
were similar to those reported for the Chios breed18 but low-
er than that reported for other Greek breeds15,20.
The LogSCC varied, however not significantly, throughout
the lactation. In general, the levels of the SCC in the milk of
the Arta breed were similar to those reported in other dairy
sheep breeds16,18. Previous studies have shown that the stage
of lactation and hand milking have an impact on SCC7,24,25.
Hand-milking, like that implemented in the present study,
has been reported to provide an efficient and uniform strip-
ping and reduce variability17.
In addition to the milk constituents reported above, average
annual milk yield for the first, second and third year was 250,
260 and 280 kg respectively with no statistical significant dif-
ferences among years. Furthermore, some physicochemical
characteristics were determined including milk density, pH
and acidity (data not shown). The average milk density was
almost identical to that reported for several Greek breeds15,20.
Average pH was higher compared to that reported in the lit-
erature16,26, but similar to that reported for the Greek breed
of Boutsiko15. Average acidity was similar to that reported by
others20,26.
It should be noted that the reasons for the conflicting find-
ings of various researchers regarding the pattern of varia-
tion of milk constituents are not clear, but it is possible that
they may be due to differences in the feeding regiments of
the ewes27, the milking procedures, the breeding or in the
climate28.
Correlations between milk
constituents
Pearson correlation coefficients among the different vari-
ables investigated are presented in Table 2. Most of the vari-
ables were correlated. Lactose was negatively correlated with
fat, protein and LogSCC and positively correlated with SNF.
These findings are in accordance with the findings of Simos
et al.20 and Sinapis24. There was a negative correlation be-
tween LogSCC and SNF which agrees with the findings re-
ported by others29,30,31. Fat was positively correlated with pro-
tein and negatively correlated with lactose.
The present study revealed that overall the milk composi-
tion of the Arta dairy sheep breed is good and within the
range reported in the literature. Since milk composition is
a key factor for milk quality and in turn affects quality of
produced dairy products, preservation or further improve-
ment of milk quality may support farmers’ income in less
favoured, rural areas.
Under this context, several other factors may affect
farmer’s income and activity in less favoured rural areas
that need to be taken into account. More specifically, under
recent changes in the Common Agricultural Policy (CAP),
low-quality land areas which have traditionally been used
as grazing lands are not eligible anymore for European
Union income support payments, thus farmers’ income
may be negatively affected32. In order to increase endurance
of rural human population in less favoured areas, under-
take of other economic activities should take place. The
most promising one is on-farm cheese manufacturing of
locally made dairy products. Production of local tradition-
al dairy products, like cheeses, is an expression of people’s
culture and lifestyle, and a reflection of the history of the
local area because traditional cheese manufacture is a re-
sult of accumulated empirical knowledge passed from gen-
eration to generation33,34. The scheme of production of lo-
cal traditional cheeses can be further supported by the in-
terest in traditional and regional foods that has become a
new consumer trend in food markets all around the world
and by the European legislation that aims to protect agri-
cultural products with geographical indication and desig-
nation of origin36.
CONCLUSIONS
The present study examined selected milk quality parame-
ters of a Mediterranean dairy sheep breed that is usually
reared in less favoured, rural and mountainous areas and re-
vealed that the milk composition of the Arta dairy sheep
breed is good. In order to support farmers’ income the good
milk quality should be maintained and other economic ac-
tivities, like on- farm cheese manufacturing should be un-
dertaken.
Level of statistical significance of each of the pairwise correlations,
*P≤0.05; **P≤0.01; *** P≤0.001
Table 2
- Correlations between the examined physicochemical
properties of Arta’s sheep milk.
Fat Protein Lactose SNF
4 BONTINIS_imp_ok 03/10/17 16:17 Pagina 187
188 Monitoring of selected milk quality parameters during lactation of a Mediterranean dairy sheep breed
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PUBBLICAZIONE ARTICOLI LARGE ANIMAL REVIEW
I medici veterinari interessati alla pubblicazione di articoli scientifici
sulla rivista “LARGE ANIMAL REVIEW” devono seguire le indicazioni
contenute nel file Istruzioni per gli autori consultabili al sito
http://www.sivarnet.it
INFORMAZIONI:
Segreteria di Redazione - redazionelar@sivarnet.it
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4 BONTINIS_imp_ok 03/10/17 16:17 Pagina 189
1998 • 2018
20° CONGRESSO
INTERNAZIONALE
16-18 MAGGIO 2018
Cremona, Centro Studi SIVAR
SIVAR - SOCIETÀ ITALIANA VETERINARI PER ANIMALI DA REDDITO
4 BONTINIS_imp_ok 03/10/17 16:17 Pagina 190
F. CECCHI, C. RUSSO, C. CANTILE, G. PREZIUSO
Department of Veterinary Science, University of Pisa, Viale delle Piagge 2, 56124 Pisa, Italy
F. Cecchi et al. Large Animal Review 2017; 23: 191-193 191
Corresponding author:
Francesca Cecchi (francesca.cecchi@unipi.it)
SUMMARY
For the development of an appropriate programme for conservation of animal genetic resources, genetic typifying is consid-
ered an important preliminary step. In this paper, we have done a preliminary genetic variability analysis of 48 adult Garfagn-
ina goats belonging to a single flock of Tuscany using 12 STR markers (MAF065, SRCRSP05, INRA023, McM527, CSRD247,
SRCRSP23, OarFCB20, TGLA53, INRA005, INRA063, ETH10, ILSTS87) some of which belonged to a markers panel validat-
ed by the International Society of Animal Genetics (ISAG) and others routinely used by the facilities of the Laboratorio di Ge-
netica e Servizi (Associazione Italiana Allevatori, Migliaro, Italy). Garfagnina is an Italian native goat breed registered on the
Tuscan regional repertory of genetic resources at risk of extinction and have a total of about 745 animals belonging to 17 flocks.
Garfagnina breed is important for livestock biodiversity preservation, being a key animal for specialized cheese market in the
Tuscan region.
For each marker the following parameters were computed: number of alleles, effective allele size, observed heterozygosity and
polymorphism information content (PIC). Allelic frequencies were estimated by direct counting. To analyze the genetic vari-
ability of the population, the following parameters were computed at population level: molecular co-ancestry coefficients (fij),
kinship distance (Dk), and inbreeding coefficient (Fi). Moreover, genetic similarities (GS) among all animals were investigat-
ed using the Individual Multilocus Genotype. The number of alleles ranged from 3 to 9 (mean 5.92) whereas the expected het-
erozygosity ranged from 0.48 to 0.83 (mean 0.69). There was a high genetic similarity within the whole population (0.43)
showing the great homogeneity of the sampled animals, as confirmed also by the small kinship distance (0.34). However in-
breeding coefficient was low (0.32). The results of this research indicate that, despite the fact that animals are considered to be-
long to the same breeding, the genetic variability of this Garfagnina goat population is acceptable for a population with a re-
duced numerical value.
KEY WORDS
Genetic variability, STR markers, Garfagnina goats.
INTRODUCTION
Italy has a long history of goats breeding and, despite a dra-
matic number contraction occurred in the last century, goat
farming is still an important reality on the Italian livestock
panorama. Many different goats breeds are diffused through-
out the entire Italian territory from the green Alpine regions
to the dry southern and island ones and they may represent a
unique source of genetic diversity. The knowledge of the ge-
netic variability is essential to preserve and exploit biodiver-
sity; the genetic variability of a population can be estimated from
genealogical data or using the short tandem repeat (STR) mo-
lecular markers both in livestock1,2,3,4,5 and in pet animals6,7. At
the molecular level more recently SNPs have also been intro-
duced8. In this context, the purpose of this work was that to
make a preliminary genetic variability analysis of the native
Garfagnina goats based on microsatellite polymorphism.
Garfagnina breed is important for livestock biodiversity
preservation, being a key animal for specialized cheese market
in the Tuscan region. Garfagnina is an Italian native goat pop-
ulation registered on the Tuscan regional repertory of genet-
ic resources at risk of extinction, with about 745 animals be-
longing to 17 flocks. The origin of this population is still un-
certain, even if it seems to derive from crossings between na-
tive goats from Alpine Arc and from the Tuscan-Emilian Apen-
nines; local breeders refer that the population was reared for
generations for its milk and meat production.
MATERIALS AND METHODS
The study was performed in a Garfagnina goat breed flock con-
sisting of 269 females and 20 males. Age ranged from 2 to 9 years.
All animals were registered in the herdbook, but genealogical
information was not available. The flock was located in the
Garfagnana district (Media Valle del Serchio, Lucca, Italy) and
a semi-extensive farming was practiced. The goats grazed dur-
ing the morning (feed supplements are given mainly over the
winter), and were housed overnight, when they received an in-
tegration of forage and feed. Flock management was of a fam-
ily farm type. Milking was practiced twice a day using a trol-
ley milking and the milk was conveyed in refrigerated tanks.
Blood samples from the 48 Garfagnina goats, were collected ac-
cording to the recommendations of the European Council
(1986) concerning animal care. Whole blood was collected in
Vacutainer tubes with K-EDTA as anticoagulant and stored at
−20 °C until genomic DNA was extracted using Qiagen QIAamp
DNA blood mini/midi kit (Qiagen, San Diego, CA, USA).
Twelve microsatellites (MAF065, SRCRSP05, INRA023,
McM527, CSRD247, SRCRSP23, OarFCB20, TGLA53, INRA005,
j
Preliminary genetic variability analysis
of the native Garfagnina goats based
on microsatellite polymorphism
j
5 Cecchi imp_OK autore_ok 03/10/17 16:21 Pagina 191
SRCRSP23 6-FAM 69-111 unknown 9 5,35 0.831 79.31%
OarFCB20 PET 86-118 2 4 2.57 0.611 56.30%
MAF065 VIC 115-151 15 9 5.87 0.830 80.97%
ILSTS87 6-FAM 137-151 28 7 5.38 0.814 78.77%
INRA005 NED 110-126 12 5 2.49 0.599 53.34%
TGLA53 PET 130-160 16 5 3.14 0.681 61.89%
McM527 NED 162-178 5 6 3.58 0.721 67.30%
SRCRSP05 VIC 153-181 21 6 4.17 0.760 72.51%
INRA063 6-FAM 169-179 18 4 1.93 0.482 40.72%
INRA023 VIC 190-220 3 7 2.91 0.656 61.98%
ETH10 NED 198-286 5 3 2.53 0.605 53.60%
CSRD247 6-FAM 211-263 14 6 3.62 0.724 67.31%
population are summarized in Table 1. In total, 71 alleles were
observed for the 12 microsatellite loci analyzed. All 12 mi-
crosatellite markers resulted to be polymorphic. Table 2 reports
the percentage of each of the most frequent alleles (>10%) for
each marker.
The most polymorphic loci were: MAF065 and SRCRSP23 (9
alleles) (Table 1) but the alleles with a frequency higher than
10% were respectively 3 and 5 (Table 2); on the contrary the
less polymorphic loci were: ETH10 (3 alleles), INRA063 and
OarFCB20 (4 alleles). All the alleles of ETH10 marker and all
the alleles of OarFCB20 had a frequency higher than 10%
(Table 2).
The PIC per locus showed only one marker with values under
the 50% (INRA063) and an average value of 64.5% (±12.30).
Genetic similarity within the population (GS), the mean mo-
lecular co-ancestry (fij), the kinship distance (Dk) and the in-
breeding coefficient (Fi) were 0.430, 0.308, 0.304 and 0.318 re-
spectively (Table 3).
INRA063, ETH10, ILSTS87), located in 12 chromosomes and
amplified in one multiplex PCR reactions, were investigated. De-
tailed information of these markers is reported in Table 1. Some
STR belonged to a markers panel validated by the International
Society of Animal Genetics (ISAG) and others were routinely
used by the facilities of the Laboratorio di Genetica e Servizi (As-
sociazione Italiana Allevatori, Migliaro, Italy).
For each marker the following parameters were computed us-
ing the Molkin v2.0 program9: number of alleles, effective al-
lele size, observed heterozygosity and polymorphism infor-
mation content (PIC). Allelic frequencies were estimated by di-
rect counting. To analyze the genetic variability of the popu-
lation, the following parameters were computed at population
level by using the MolKin program9: molecular co-ancestry co-
efficients (fij), kinship distance (Dk), and inbreeding coefficient
(Fi). The molecular co-ancestry between 2 individuals, i and
j (fij), is the probability that two randomly sampled alleles from
the same locus in 2 individuals are identical by state (Caballero
and Toro, 2002). The molecular co-ancestry of an in-
dividual i with itself is self-co-ancestry (si), which is
related to the coefficient of inbreeding of an individual
i (Fi) by the formula Fi = 2si -1. In turn, the kinship
distance (Dk) between 2 individuals i and j is Dk =
[(si +sj)/2] - fij10. MolKin computes within-breed mo-
lecular co-ancestry and Dk by simply averaging the
corresponding values for all the within-population
pairs of individuals. Moreover, genetic similarities
among all animals were investigated by comparing the
individual multilocus genotype of each individual. Ge-
netic similarity is defined as P=A/2L, where P is the
proportion of common alleles (A) in relation to the
2L possibilities (L=number of considered loci). The
similarities between each pair of individuals were then
averaged over the whole population.
RESULTS
The results of the microsatellite analysis in term of
number of alleles observed, alleles size, PIC and ob-
served heterozygosity of the analyzed Garfagnina goat
192 Preliminary genetic variability analysis of the native Garfagnina goats based on microsatellite polymorphism
Marker OarFCB20 Marker ILSTS87
Allele 93 95 97 99 Allele 137 139 141 143 145
Frequency 0.23 0.13 0.53 0.11 Frequency 0.17 0.16 0.26 0.17 0.19
Marker INRA005 Marker MAF065
Allele 115 117 Allele 121 131 133
Frequency 0.54 0.31 Frequency 0.15 0.28 0.22
Marker McM527 Marker TGLA53
Allele 152 162 164 Allele 134 136 146
Frequency 0.24 0.26 0.39 Frequency 0.36 0.38 0.21
Marker INRA063 Marker SRCRSP05
Allele 173 175 Allele 163 171 173 179
Frequency 0.29 0.66 Frequency 0.26 0.10 0.36 0.18
Marker ETH10 Marker INRA023
Allele 203 205 207 Allele 197 211 215
Frequency 0.21 0.48 0.31 Frequency 0.15 0.53 0.19
Marker CSRD247 Marker SRCRSP23
Allele 228 230 240 Allele 79 91 95 101 105
Frequency 0.30 0.26 0.33 Frequency 0.34 0.14 0.10 0.11 0.14
Table 2
- Percentage of each of the most frequent alleles (> 10%) for each
marker.
Table 1
- Locus, Dye, range, number of alleles, effective allele size (EfAlSize), observed heterozygosity (Ho) and polymorphism information
content (PIC), for the 12 microsatellite loci.
Locus Dye Range Chromosome N° alleles Ef.AlSize Ho PIC
5 Cecchi imp_OK autore_ok 03/10/17 16:21 Pagina 192
tionship between the animals observed by the mi-
crosatellites analysis.
In conclusion, the analysis performed with the use
of 12 microsatellite markers showed that despite all
goats originated from the same flock and are pre-
sumably subdivided into 3 groups with different as-
cenders, the inbreeding was lower than that report-
ed in all the paper previously mentioned and the ge-
netic variability was acceptable for a population with
a reduced effective numerical value. Future work may include
replication of this study with a larger number of animals be-
longing to different flocks.
Acknowledgements
This work was supported by grants of the University of Pisa
(PRA 2016 and Fondi Ateneo).
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Ho*: observed heterozygosity. fij#: molecular co-ancestry coefficients.
Fi‡: inbreeding coefficient. Dk§: kinship distance. GSll: genetic similarity.
F. Cecchi et al. Large Animal Review 2017; 23: 191-193 193
Total 48 5.92±1.881 0.693 0.308 0.318 0.340 0.430±0.103
Table 3
- Within-population diversity.
NAverage n° Ho* fij#Fi‡Dk§GSll
of alleles
DISCUSSION AND CONCLUSION
There is dearth of published reports on the genetic variabili-
ty and on the number of alleles, their size and frequencies for
microsatellite loci in goats. Although a comparison with oth-
er breeds can be biased due to the different marker sets used
by different authors, it may be noted how the mean number
of alleles per locus was slightly lower than that reported by Ra-
mamoorthi et al.11 on the Barbari goats, and by Sechi et al.12
on three Sardinian goat populations, but similar to what ob-
served in Orobica and Girgentana goats by Negrini et al.4. Oro-
bica and Girgentana exhibited small genetic variability and
therefore evidence of a recent bottleneck. However, our ani-
mals derived from a single flock.
The PIC estimated in the present study is comparable with that
obtained in other goat breeds, such as Saanen4. The PIC was
originally introduced by Botstein et al.13. It refers to the value
of a marker for detecting polymorphism within a population,
depending on the number of detectable alleles and the distri-
bution of their frequency and has been proved to be a gener-
al measure of how informative a marker is; the higher is the PIC
value, the more informative a marker is. In the present study,
MAF065 and SCRRSP23 and ILSTS87 microsatellites ap-
peared the most informative, whereas INRA063 the less in-
formative.
Ten of the 12 markers considered in this research were used also
by Sechi et al.12 for the study of the genetic variability in Mal-
tese, Sardo autochthonous goats and their mixed blood pop-
ulation, and by Negrini et al.4who analyzed the genetic struc-
ture of eight Italian goat breeds (Camosciata delle Alpi, Valo-
dostana, Bionda dell’Adamello, Orobica, Grigia Molisana, Gir-
gentana, Argentata dell’Etna, and Sarda).
Genetic similarity within our population is higher than reported
in other species such as cattle3and sheep2,5, but lower than that
reported in two Italian autochthonous donkey genetic types14
and in Bracco Italiano dog breed6. With the exception of the val-
ues reported in Amiata and Viterbese donkey breeds 14 and in
Bracco Italiano dog breed6, the values observed in our study re-
garding the mean molecular co-ancestry was clearly greater than
those reported in sheep2,5 and in cattle3. Conversely, the kinship
distance was smaller than that reported in literature.
In a previous study15 the same animals were also genotyped us-
ing the Illumina GoatSNP60 BeadChip (Illumina Inc., San
Diego, CA), containing 53,347 SNPs, designed by the Inter-
national Goat Genome Consortium (IGGC). Selected SNPs were
used to verify the population structure of this goat population.
A matrix of genomic relationships was calculated and a ma-
trix of genetic distances was built. Three main clusters (Group
1: 33 animals; Group 2: 6 animals; Group 3: 9 animals) has been
identified. Even if it was not possible to trace back the ances-
tors of any individual, we postulated that each cluster refers to
daughters of related individuals. This explains the high rela-
5 Cecchi imp_OK autore_ok 03/10/17 16:21 Pagina 193
5 Cecchi imp_OK autore_ok 03/10/17 16:21 Pagina 194
Garantire il benessere nell’allevamento
dell’asino per la produzione di latte:
un’analisi della legislazione
F. SPADOLA, G.L. COSTA, M. MORICI, C. INTERLANDI, B. NASTASI, M. MUSICÒ
Dipartimento di Scienze Veterinarie, Università degli Studi di Messina
F. Spadola et al. Large Animal Review 2017; 23: 195-197 195
Autologous prosthesis for the surgery
of two simultaneous hernias in a calf
N
Autore per la corrispondenza:
Marcello Musicò (marmus@unime.it).
SUMMARY
The authors propose a rare case of double hernia in a female, red freisian calf, seventeen months old. Hernia that occurs most
frequently in calf is related to hereditary factors or mainly traumatic causes. The clinical examination of the lesion led to de-
tect two similar lesions, round and smooth, not-hot and not-sore, the first localized to the umbilical region, and the second,
more caudal and larger, in a paramedian position. Both lesions were palpable in the abdomen through regular continuous and
non-communicating solutions. After some basic analysis, the calf received general anaesthesia and underwent standard sur-
gery, but with the application of autologous prosthetic material, seen the location and peculiarity of the lesion. After two
months the calf was sound and all her functions restored. Double hernia requires to strengthen the suture with prosthetic ma-
terial. A flap of the hernial bag wall has been used as prosthetic material, since is strong and well-vascularized, provides excel-
lent tissue compatibility and guarantees that, thanks to constant blood flow, there is always the presence of cellular reaction el-
ements with the formation of a stiff fibrin cloth and a healthy healing tissue. After the surgical procedures, the calf was awak-
ened regularly. At a first check the day after the surgical procedure, the animal regularly resumed eating and drinking. After
two months, the patient’s follow-up confirmed the complete healing of the lesions, with functional recovery of the herniated
organs and the general condition of the subject. The use of an autologous prosthesis comes from a twofold consideration: the
great stress that tissues are subjected to an animal with a double hernia, requires the application of a reinforcement that re-
mains stable for at least 15 days in the postoperative. In addition, the autologous prosthesis, made up of well-vascular tissue,
has excellent compatibility between tissues. Such features favor the formation of new granulation and fibrin tissues in a short
time, thus reducing healing times.
KEY WORDS
Calf, hernia, autologous prosthesis, laparocele.
INTRODUCTION
Hernia that occurs most frequently in calf is related to hered-
itary factors or mainly traumatic causes. These are mainly
external hernias affecting the abdominal region, the inguinal
region and the pelvic region1,2,4. The hernia content may
sometimes reach a considerable size, with the loss of the pos-
sibility of reduction2,4,6. In acute cases, as in congenital cases,
the lesion begins without significant systemic involvement.
In chronic cases, or ones complicated by consecutive or in-
flammatory lesions (umbilical hernia above all), the anam-
nesis reports a progressive deterioration of the organic con-
ditions, with decrease in growth and / or milk production1,2,4.
The lesion usually detects a variable volume swelling, gener-
ally rounded and regular, not hot and not painful. An accu-
rate compression of the hernia, in most cases, allows the dis-
appearance of the swelling and perceiving the rupture
door1,2,4,9. In chronic cases, where phenomena of flogosis and
sclerosis modify the anatomical relationships between the
various structures, it is still possible to palpate the lesion
without the animal’s painful reactions, but it is impossible to
reduce swelling. In complicated cases of secondary injuries,
in addition to a general state of general suffering, the lesion
is hot, very sore and intractable6,7,8. For differential diagnosis,
it is sufficient to perform a puncture of the lesion. In some
cases, however, the above method is useless: for example, in
some chronicized umbilical abscesses, the walls are so thick,
often concatenated, and the pus is so dense that the centesis
does not provide significant data. In these cases, an ultra-
sound examination should be performed to obtain verifiable
data on the tumor content and, therefore, to establish appro-
priate therapy6,7,8.
There are various techniques in the literature that are use-
ful for the treatment of hernia: the size of the lesion should
be taken into account7,9,10. It should be remembered that
subjects with congenital hernia should be excluded from
reproduction as it is a transmissible hereditary factor. By
leaving behind the conservative techniques described in the
literature, it is necessary to state that larger ruptures, as well
as the complicated ones, require classical surgical thera-
py7,8,9. If there is a doubt about a poor suture holding, for
example in very large animals or if the abdominal walls are
altered by the chronicization of the lesion, some authors
suggest that a network of resorbable synthetic material be
applied to the suture of the rupture door, in order to rein-
force the latter6,9,10. In this regard, since the value of these
alloprostheses is recognized in the success of surgery and in
Spadola_imp_ok 03/10/17 16:22 Pagina 195
196 Autologous prosthesis for the surgery of two simultaneous hernias in a calf
the prevention of recurrences, although their costs are high
and certainly unrepresentable in the case of income-pro-
ducing animal surgery, it is thought to replace synthetic
prosthetic material with an autologous prosthesis, obtained
by creating a vascularized flap from the hernia bag6,9,10. This
surgical method is intended to confer considerable resist-
ance to erniorraphy and to prevent recurrences, without
using synthetic prosthetic materials.
MATERIALS AND METHODS
A seventeen months old female calf, of red-headed Italian,
was enrolled in this study. It was visited and operated at the
same time as a field clinic, carried out in the province of Ra-
gusa (Sicily).
The objective examination of the lesion allowed to detect a
swelling of size 8x7 cm, of round shape and regular, not hot
and not painful, localized to the umbilical region, and a sec-
ond caudal and larger, of the size of 12x9 cm, in a paramedi-
an position, with the same anatomical characteristics as the
first one.
The lesion doors had a diameter of 4 cm and 7 cm respec-
tively (Fig. 1). Both swellings were palpable in the abdomen
through regular, round, non-communicating lesion doors
(Fig. 2). Upon completion of the clinical trial, a diagnosis of
double laparocele was formulated. Therefore, it has been
programmed for surgery for a few days. The subject was
placed on 24-hour pre-operative fasting, while the water was
suspended from the previous evening. Before the operation,
the animal was subjected to a further accurate clinical and
anesthesiological examination. Haematological and haema-
tochemicalanalysis have been performed that have given val-
ues to the standard.
The patient was anesthetized with xylazine hydrochloride,
equal to 0.2 mgkg-1, followed by an intravenous ketamine
hydrochloride infusion, at a dose of 5 mgkg-1, then tramadol
1 mgkg-1, administered at the end of the dorsal decubitus3,5.
Throughout the duration of the intervention, intravenous
access was maintained by NaCl 0.9% perfusion. The surgery
site, that is skin, subcutaneous tissue, surrounding tissues
and hernial door, has been infiltrated, with full thickness,
with lidocaine at 2%3,5.
Once reached an appropriate stage of anesthesia and anal-
gesia, after having prepared the surgical field according to
surgical art, the skin was incised and then, with the scissors,
bluntly, the surrounding tissues, to isolate the hernia bag.
This was incised, thus highlighting the hinged door and its
contents.
The dislocated organs were appropriately repositioned
through the hinged door. After having gunned the sclerotic
edge of the door, traditional erniorraphy was performed,
with detachable nodous points and monofilament n. 2 ab-
sorbable wire. The remaining hernia bag was shaped and
used, by attachment and fixation, to reinforce the suture,
with detached nipple stitches, also with resinable monofila-
ment n. 2 (Fig. 3). It was then sutured the subcutaneous
plane with detached sutures and the skin.
The interventions carried out on the two lesions overlap as
surgical procedures: first, on the onphalocele and then on the
other lesion, with two separate operating fields but with the
same surgical technique.
RESULTS
Immediately after the surgical procedures, the calf was
awakened regularly. At a first check the day after the surgi-
cal procedure, the animal regularly resumed eating and
Figure 1
- Seventeen months old female calf. It is possible to see
the double swelling located at the level of the caudal abdominal
wall.
Figure 2
- The openings of the double hernias. With the simple fin-
ger pressure, it is possible to replace the herniated organs.
Figure 3
- View of the surgical field, with the autologous flap al-
ready sutured in place.
Spadola_imp_ok 03/10/17 16:22 Pagina 196
F. Spadola et al. Large Animal Review 2017; 23: 195-197 197
drinking. After two months, the patient’s follow-up con-
firmed the complete healing of the lesions, with functional
recovery of the herniated organs and the general condition
of the subject.
DISCUSSIONS AND CONCLUSIONS
Although the external appearance led immediately to think
of two congenital hernias, intraoperatively, by the evaluation
of the rupture’s rings ring, it was suggested that paramedic
hernia could be of traumatic nature, occurring during child-
birth or immediately postpartum. In fact, this second hinged
door was presented with jagged and uneven margins, unlike
the umbilical, regular, and fibrous margins.
The idea of using an autologous prosthesis came from a
twofold consideration: in the meantime, the great stress to
which the tissues are subjected, especially in an animal with a
double hernia, which requires the application of a reinforce-
ment that remains stable for at least fifteen days in postopera-
tive; in addition, the hernia bag, strong and well-vascularized,
provides excellent tissue compatibility that, thanks to constant
blood flow, guarantees the presence of cellular defense and re-
active elements with the formation of a stiff fibrin layer.
References
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Large Animal Review 2017; 23 199
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200
LA RASSEGNA DI VET-JOURNAL
Rubrica a cura di Maria Grazia Monzeglio
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http://www.evsrl.it/vet.journal/
• Notizie scientifiche
Archivio bibliografico • Servizio Cytovet
Influenza D: un’infezione emergente
nei suini italiani
Rilevanza epidemiologica del patogeno
in aumento, secondo uno studio condotto
nella Valle del Po
Il virus dell’Influenza D (IDV), nuovo membro del-
la famiglia Orthomyxoviridae, è stato segnalato per
la prima volta nei suini in Oklahoma nel 2011 e suc-
cessivamente riscontrato nei bovini in Nordamerica
ed Eurasia. Al fine di indagare la circolazione di IDV
nei suini iitaliani, nel periodo tra giugno 2015 e
maggio 2016 si effettuavano test biomolecolari e vi-
rologici su 845 campioni clinici prelevati da 448 al-
levamenti suini affetti da problemi respiratori e lo-
calizzati nella valle del Po. Si effettuavano test siero-
logici su 3698 sieri suini, includendo sieri prelevati
nel 2009, così come campioni prelevati nel 2015 nel-
la stessa regione. Si identificava il genoma virale in
21 campioni (2,3%) di 9 allevamenti (2%), mentre il
virus veniva isolato con successo da tre campioni. Le
analisi genetiche evidenziavano che gli IDV dei sui-
ni italiani erano strettamente correlati al cluster
D/swine/Oklahoma/1334/2011. I sieri prelevati nel
2015 mostravano un’elevata prevalenza di titoli an-
ticorpali IDV (11,7%), mentre i sieri del 2009 mo-
stravano tassi di positività significativamente infe-
riori (0,6%).
I risultati indicano che la rilevanza epidemiologica
del patogeno è in aumento e che sono necessari ul-
teriori indagini per comprendere la patogenesi,
l’epidemiologia e il possibile potenziale zoonosico di
questo virus emergente, concludono gli autori.
“Influenza D in Italy: towards a better understanding of an
emerging viral infection in swine.” Foni E, Chiapponi C, Ba-
ioni L, Zanni I, Merenda M, Rosignoli C, Kyriakis CS, Luini
MV, Mandola ML, Bolzoni L, Nigrelli AD, Faccini S. Sci Rep.
2017 Sep 15; 7 (1): 11660.
Patologie neurologiche dei ruminanti:
studio retrospettivo di coorte
Paresi spastica, osteomielite vertebrale
e listeriosi le diagnosi più frequenti nel corso
di 10 anni
Nel corso di 10 anni (2006-2016), presso l’Universi-
tà di Glascow in Scozia si osservavano 96 ruminanti
con segni neurologici, costituenti il 5,4% di tutte le
visite effettuate. Erano descritti 47 diversi segni neu-
rologici di presentazione e il 79% dei pazienti pre-
sentava anomalie di andatura. Tutti i casi con ano-
malie in più di 4 su 10 categorie neurologiche dece-
devano o erano soppressi per motivi di benessere. I
vitelli avevano una probabilità significativamente
maggiore di presentare patologie neurologiche ri-
rella pyogenes erano localizzati all’interno dell’endo-
metrio, sulla superficie endometriale e nello stroma
caruncolare, ma la loro presenza non era associata a
infiammazione. I dati del sequenziamento masssivo
parallelo del gene del 16S rRNA di un sottogruppo di
15 bovine indicavano che i batteri più abbondanti
erano della famiglia Porphyromonadaceae, seguita
da Ruminococcaceae e Lachnospiraceae. I risultati
indicano che l’utero bovino non è un ambiente steri-
le durante la gravidanza, come precedentemente rite-
nuto, e che la bovina può condurre la gravidanza an-
che in presenza in utero di alcuni batteri potenzial-
mente patogeni, concludono gli autori.
“Presence of bacteria in the endometrium and placentomes of
pregnant cows”. Cecilia Christensen Karstrup. Theriogenolo-
gy. September 1, 2017. Volume 99, Pages 41-47.
Anastomosi vescicoprepuziale per
l’urolitiasi ostruttiva del becco
Una procedura chirurgica praticabile per
i casi ricorrenti, con esito clinico favorevole
in uno studio preliminare
Uno studio descrive una nuova tecnica chirurgica
per la correzione dell’urolitiasi ostruttiva ricorrente
nel becco. Si rivedevano le cartelle cliniche di 4 bec-
chi castrati sottoposti ad anastomosi vescicoprepu-
ziale (VPA) per il trattamento di questa condizione.
Tutti gli animali erano stati sottoposti ad almeno
una procedura chirurgica (mediana: 2,5; range: 2-4)
per la correzione dell’urolitiasi, prima di essere sot-
toposti a VPA. Le complicazioni postoperatorie in-
cludevano la rimozione prematura del catetere dalla
vescica (1 soggetto), cistite batterica (2) e formazio-
ni di ascessi (1). In un soggetto si verificava la steno-
si dell’anastomosi 3 mesi dopo la procedura iniziale
e veniva sottoposto a una seconda VPA; un soggetto
decedeva 7 mesi dopo la chirurgia a causa di una
grave idronefrosi acuta e insufficienza renale.
La sopravvivenza a lungo termine ≥12 mesi era buo-
na, con 3/4 animali (75%) o 3/5 procedure VPA
(60%) con flusso urinario non ostruito a 12 mesi.
L’anastomosi vescicoprepuziale è una procedura
chirurgica praticabile per la correzione dell’urolitia-
si ostruttiva ricorrente nel becco e può determinare
un esito clinico favorevole. Sono auspicabili ulterio-
ri studi su più ampie popolazioni caprine per la va-
lutazione della idoneità della tecnica in questa con-
dizione, concludono gli autori.
“Vesicopreputial anastomosis for the treatment of obstructive
urolithiasis in goats.” Elizabeth Erin Cypher et al. Vet Surg.
February 2017; 46 (2): 281-288.
spetto ai bovini adulti. Le lesioni erano più spesso
localizzate al midollo spinale nel 47% delle pecore
(16), al sistema nervoso periferico nel 45% dei bovi-
ni (28) e a livello cerebrale nel 41% della popolazio-
ne complessiva (39). Le cause più comuni delle pa-
tologie neurologiche osservate erano infettive o in-
fiammatorie (28%, 27). Una diagnosi definitiva era
raggiunta nell’84% (81) dei pazienti. Quando era di-
sponibile un esame autoptico, esso produceva una
diagnosi nel 70% (50) dei casi e contraddiceva la
diagnosi clinica nel 38% dei casi (26).
Le diagnosi più frequenti nei ruminanti nel corso di
10 anni erano paresi spastica, osteomielite vertebra-
le e listeriosi.
“Ruminant neurological disease: a retrospective cohort study”.
Giles L, et al. Vet Rec. 2017 Sep 5.[Epub ahead of print].
L’utero bovino gravido non è
un ambiente sterile
Batteri presenti nel 90% circa dei campioni
endometriali e placentari di uno studio,
non associati a infiammazione
Nella maggior parte delle bovine si verifica un’inva-
sione batterica dell’utero durante il periodo po-
stpartum ma si ritiene generalmente che tali batteri
siano eliminati prima della gravidanza successiva.
L’utero gravido è quindi stato fino ad oggi conside-
rato un ambiente sterile, tuttavia questo assunto è
stato ora messo in discussione da recenti studi nel-
l’uomo che indicano che è possibile la presenza di
batteri nella placenta della gravidanza a termine sen-
za associazione con l’aborto.
Uno studio ha indagato se fossero presenti batteri
nell’utero gravido bovino. Si prelevavano campioni
di endometrio inter-caruncolare e di placentoma da
43 bovine gravide al macello, sottoponendoli a esame
istopatologico, ibridazione fluorescente in situ e se-
quenziamento massivo parallelo. Si osservavano bat-
teri nei tessuti del 90,7% (39/43) delle bovine me-
diante ibridazione fluorescente in situ. Fusobacte-
rium necrophorum, Porphyromonas levii e Truepe-
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