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Source publication
The present study was planned to detect the genetic elements of Mannheimia haemolytica and Pasteurella multocida in pneumonic sheep lungs. Pneumonia was diagnosed on the basis of gross pathological lesions. Lung tissues were collected at necropsy of sheep (n = 96) and subjected to isolation of total DNA. The M. haemolytica-specific PHSSA and Rpt2 g...
Context in source publication
Context 1
... postmortem examination showed congestion and petechial and ecchymotic type of hemorrhages in the lung tissues of dead sheep (Figures 1 and 2). Patchy to diffuse, reddish-brown to greyish-red areas of consolidation and hepatization of lungs was predominantly recorded on gross pathological examination. ...
Citations
... Similar to the current investigation, previous reports have described M. haemolytica and P. multocida isolation and identification on the basis of culture method and biochemical features, as well as PCR tests addressing the particular genes. In addition, according to Singh et al. [34], Singh et al. [35] and Hassan et al. [36], the exact molecular detection of P. multocida using 16S rRNA and the KMT1 gene has a substantial effect on the epidemiology and management of pasteurellosis in small ruminants. In addition, it is asserted that molecular typing techniques enable quick identification of bacteria and have been shown to be more accurate than culture and biochemical testing [19,37]. ...
Objective
This research aimed to assess the prevalence of caprine pasteurellosis, isolate and identify pasteurellosis (Mannheimia haemolytica and Pasteurella multocida) in pneumonic goats, and discover the main bacterial cause of pneumonia.
Materials and Methods
One hundred and five samples (94 nasal swabs and 11 lung tissues) from goats suspected of having pneumonia were taken and transferred aseptically to the laboratory. Following the processing of the collected samples, Pasteurella spp. was isolated with the aid of plate culture methods. Biochemical characteristics were used to identify all bacterial isolates, which were then verified by polymerase chain reaction (PCR). Antimicrobial susceptibility testing was also carried out to evaluate the sensitivity profiles of various antibiotics. The Pasteurella haemolytica serotype-specific antigen (PHSSA) gene was used to identify isolates of M. haemolytica, and the KMT1 gene was used to identify isolates of P. multocida.
Results
From the 105 clinically suspicious samples, 51 (48.57%) were identified to be Pasteurella spp. through bacteriological testing and also by PCR targeting the 16S rRNA gene. Of these, 47.87% (45/94) were nasal swabs, and 54.55% (6/11) were lung tissues. Among confirmed samples, 70.59% (36/51) were identified as M. haemolytica, and 29.41% (15/51) were identified as P. multocida. Resistance to tetracycline, streptomycin, oxytetracycline, gentamicin, and ceftriaxone was found in 50%–83% of the isolates. In addition, PCR identified the PHSSA and KMT1 genes from isolates of P. multocida and M. haemolytica, respectively.
Conclusion
The present study revealed that M. haemolytica and P. multocida primarily caused pasteurellosis in pneumonic goats in Bangladesh. However, when treating these animals, the proper choice of antimicrobials should be made to control this disease.
... Of the 213 cultures two (2) nasal swabs were tentatively identi ed as M.haemolytica. This nding is lower than the ndings of [32], [33], [34] whose reports were 2.22% in sheep and goat, 11.2% in sheep and 14.10% and 11.80% in sheep and goats respectively. ...
Background:Mannhemia haemolytica is one of the most important bacteria among causative agent of pneumonic pasteurellosis in small ruminants throughout worldwide and it is also economically devastating pathogen in Ethiopia.
Methods: A cross sectional study was carried out from November 2021 to May 2022 with the aim to identify Mannheimia haemolytica from sheep and goat in Sebeta and Holeta town, Oromia special zone, Ethiopia. A total of 235 samples (213 nasal swabs and 22 whole blood) were collected. Sheep and goat with clinical signs suggestive of pneumonic pasteurollosis were purposively sampling. Bacterial identification was conducted using biochemical, Biolog, Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI TOF MS) and Real time PCR detection. Moreover, antimicrobials susceptibility test was also conducted on the identified bacterial isolates using disc diffusion method.
Results: The result showed that from a total of 235 samples, only two nasal swab samples were positive for M. haemolytica (0.85%). The two isolates were confirmed by all the tests and similar result was obtained by; biochemical, Biolog, MALDI TOF MS and real time PCR. Up on antimicrobial susceptibility testing, the two isolates were resistant to Streptomycin, Erythromycin and Clindamycin whereas they were susceptible to Tetracycline, Chloramphenicol, Trimethoprim/sulfonamides and Penicillin. Generally, this study revealed that M. haemolytica is among the causative agent of pneumonic pasteurellosis in sheep and goat in the study area. Although, the other remaining bacteria responsible for the disease.
Conclusion: The research suggests that a combination of diagnostic methods such as MALDI TOF MS, Biolog, and real-time PCR should be used, as well as for a more in-depth investigation to identify the strain or serotype of M. haemolytica using advanced molecular sequencing and also analysis of the remaining causal agent from various species and locations in countries are significant to address the present vaccination and antibiotic resistance issues.
... P. multocida is a well-known cause of pneumonic pasteurellosis in small ruminants (Assefa and Kelkay, 2018;Cid et al., 2020;Singh et al., 2018;Odugbo et al., 2006;Sarangi et al., 2015;Rawat et al., 2019); however, few studies have investigated its prevalence in pneumonic sheep (Lindström et al., 2018;Singh et al., 2018;Odugbo et al., 2004;Marru et al., 2013). In goats, studies on P. multocida prevalence are even scarcer and have included several hosts (Kumar et al., 2004;Shirzad Aski and Tabatabaei, 2016). ...
... P. multocida is a well-known cause of pneumonic pasteurellosis in small ruminants (Assefa and Kelkay, 2018;Cid et al., 2020;Singh et al., 2018;Odugbo et al., 2006;Sarangi et al., 2015;Rawat et al., 2019); however, few studies have investigated its prevalence in pneumonic sheep (Lindström et al., 2018;Singh et al., 2018;Odugbo et al., 2004;Marru et al., 2013). In goats, studies on P. multocida prevalence are even scarcer and have included several hosts (Kumar et al., 2004;Shirzad Aski and Tabatabaei, 2016). ...
... This study investigated the prevalence of P. multocida in a relatively large (n = 349) sample of pneumonic lungs from sheep and goats. P. multocida was isolated from 29.4 % and 13.8 % of sheep and goat pneumonic lungs, respectively, indicating that P. multocida is widely prevalent in sheep and goat farms in the province of Khuzestan (southwestern Iran), as reported for other geographic regions (Odugbo et al., 2004;Singh et al., 2018). Although a seroprevalence of 16.9 % was reported in a study of apparently healthy goats in Ethiopia (Assefa and Kelkay, 2018), there is a lack of information on the prevalence of P. multocida in pneumonic goat lungs. ...
Pasteurella multocida is an important cause of pneumonic pasteurellosis in small ruminants. Its prevalence was investigated in 349 pneumonic lungs from sheep (n = 197) and goats (n = 152), and genotypes of isolates were determined by capsular and lipopolysaccharide (LPS) typing as well as by virulotyping based on the detection of 12 virulence-associated genes. P. multocida was isolated from 29.4% of sheep lungs and 13.8% of goat lungs. A (78.5%) and D (21.5%) capsular types, as well as L3 (41.8%) and L6 (57.0%) LPS genotypes, were detected, with the A:L6 genotype being the most prevalent in both sheep (59.6%) and goat (52.4%) isolates. A total of 19 virulence profiles (VP) were detected, seven non-toxigenic and 12 toxigenic, which correlated with the capsular–LPS genotype. All isolates of each VP belonged to the same LPS and capsular genotype, except for one isolate of VP1. The diversity in VP was higher among toxigenic (0.29) than non-toxigenic (0.18) isolates. Moreover, the toxigenic VPs showed more diversity in their capsular-LPS genotypes, with the two main toxigenic VPs belonging to genotypes D:L3 (VP2) and A:L3 (VP3). Therefore, the abundance of toxigenic isolates among sheep and goat isolates does not seem to correspond to the expansion of a more virulent lineage associated with pneumonic pasteurellosis in small ruminants. The most prevalent genotypes among sheep isolates were the non-toxigenic VP1:A:L6 (41.4%) and the toxigenic VP3:A:L3 (17.2%) genotypes, whereas the most prevalent among goat isolates were the toxigenic VP2:D:L3 (33.3%) and the non-toxigenic VP1:A:L6 (14.3%) and VP4:A:L6 (14.3%) genotypes. These prevalent toxigenic and non-toxigenic genotypes seem to be epidemiologically relevant in pneumonic pasteurellosis of small ruminants.
... This overall detection rate (31%) is following some previous studies (Tilaye 2010;Alemneh and Tewodros 2016), which have found prevalence rates of 32.6 and 28.4%, respectively. However, the majority of previous studies have reported higher prevalence rates (Gilmour 1974;Aschalew 1998;Singh et al. 2018). This variation in detection rates might be due to the ecology of the study areas, seasonal variation, and the presence of predisposing factors (Abera et al. 2014). ...
AB S T RA C T Pneumonic pasteurellosis is a common and economically important type of ovine pneumonia. No previous study about the disease in Al Madinah Region, Saudi Arabia. Thus, this study was conducted to determine the association rate of Mannheimia haemolytica and Pasteurella multocida with pneumonia in sheep and to update data about their antimicrobial susceptibility pattern. A total of 100 samples (57 nasal swabs and 43 lung tissues) were collected from diseased and animals suspected to have died of pneumonia. Samples were subjected to bacteriological examination, biochemical identification of isolates by VITEK2 system, direct molecular identification by real-time PCR (RT-PCR), and antibiotic sensitivity testing of isolates. The results showed an overall detection rate of 31% for M. haemolytica (25%) and P. multocida (6%). Only 6% isolates were confirmed by VITEK 2 as M. haemolytica, with probability reached 99%. While, direct molecular method revealed that 20.2% samples were positive for M. haemolytica and 6.4% for P. multocida specific 16S rRNA genes. M. haemolytica isolates were found sensitive to oxytetracycline, nitrofurantoin, trimethoprim/sulfamethoxazole, ciprofloxacin, cefoxitin, ceftazidime, ceftriaxone, imipenem, and tigecycline, in order. While, they were found completely resistant to cloxacillin, streptomycin, and amoxicillin/clavulanic acid. In conclusion, the detection rate of M. haemolytica emphasized its role as a major cause of ovine pneumonia. Besides, our results invigorated the role of direct molecular detection and recommend it for laboratory differential diagnosis. The isolates were resistant to limited antimicrobial agents, nevertheless, the antimicrobial susceptibility test is important for proper treatment.
... The diversity of M. haemolytica capsular types was very similar in lungs of clusters 1 and 3 (eight and seven capsular types, respectively; Table 2). This diversity consolidates that there is high degree of heterogeneity in M. haemolytica serotypes colonizing the lungs of sheep (Odugbo et al., 2004;Fernández et al., 2016;Singh et al., 2018) and confirms the multiplicity of serovars capable of causing pneumonia in sheep. Despite this diversity and the wide variation in the prevalence of M. haemolytica capsular types isolated from the lungs of sheep, capsular types A6, A7, and A9 were the most commonly isolated (Table 2), a result in line with earlier studies (Odugbo et al., 2004;Fernández et al., 2016;Singh et al., 2018). ...
... This diversity consolidates that there is high degree of heterogeneity in M. haemolytica serotypes colonizing the lungs of sheep (Odugbo et al., 2004;Fernández et al., 2016;Singh et al., 2018) and confirms the multiplicity of serovars capable of causing pneumonia in sheep. Despite this diversity and the wide variation in the prevalence of M. haemolytica capsular types isolated from the lungs of sheep, capsular types A6, A7, and A9 were the most commonly isolated (Table 2), a result in line with earlier studies (Odugbo et al., 2004;Fernández et al., 2016;Singh et al., 2018). No significant differences were detected in serotype prevalence between lungs with and without lesions (data not shown). ...
... Moreover, cluster 4 lungs also showed higher frequencies of severe (20.8%) and moderate (25.0%) consolidation lesions (Table 2). These results point out that, unlike in acute clinical cases (Singh et al., 2018), in subclinical or chronic cases of pneumonic pasteurellosis such as those investigated in this study, P. multocida, compared to M. haemolytica, is expected to develop more severe pathological lesions. Coinfections with more than one pathogen usually yield significantly more severe pathological findings (Stipkovits et al., 2012;Liu et al., 2017;Sajid, 2017). ...
This study investigated the association of Pasteurella multocida isolation and the molecular characteristics of the isolates with the presence of pneumonic lesions in lambs at slaughter to assess its importance as a causative agent of pneumonic pasteurellosis compared with Mannheimia haemolytica. P. multocida was isolated from the 13.9% and 2.7%, and M. haemolytica from the 36.4% and 26.8%, of lungs with and without lesions, respectively (P < 0.05). Both microorganisms were frequently coisolated (23.2% and 12.5% from lungs with and without lesions, respectively). Isolation of P. multocida alone exhibited greater strength of association with pneumonic lesions (OR 11.4; 95% CI 3.2–40.6) than that exhibited by M. haemolytica alone (OR 3.0; 95% CI 1.6–5.4). Cluster analysis grouped the lungs into four clusters characterized by the isolation of M. haemolytica or P. multocida alone (clusters 1 and 4), coisolation of both microorganisms (cluster 3), and isolation of neither (cluster 2). Cluster 4 lungs exhibited higher frequencies of pneumonic lesions (87.5%) and severe (20.8%) and moderate (25.0%) lesions. Lungs coinfected with both pathogens (cluster 3) did not exhibit a higher frequency of severe and moderate consolidation lesions (6.1% and 14.3%, respectively), suggesting that P. multocida and M. haemolytica do not act synergically to cause more severe pneumonic infections. The greater strength of association of P. multocida isolation with pneumonic lesions together with the higher severity of the lesions caused could indicate a greater role played by this pathogen in the aetiopathogenesis of pneumonic pasteurellosis in sheep than is commonly assumed.
... Therefore, our qPCR was ten times more sensitive than the conventional PCR. The limits of detection of other PCR tests used for P. multocida have not been determined (Miflin and Blackall, 2001;Liu et al., 2004;Sabiel et al., 2012;Singh et al., 2018). The threshold limit of detection of PCR for the KMT1, HSB, PSL and Miflin primer sets were 500 pg, 50pg, 50pg and 5ng of DNA of P. multocida per assay, respectively (Jonathan and Arora, 2012). ...
Pasteurella multocida is one of the bacterial species involved in cases of ovine respiratory complex that has been implicated to cause significant economic losses in sheep production system worldwide. The present study was undertaken with the aim of evaluating a SYBR Green dye based real time PCR assay targeting KMT1 gene for the detection of P. multocida. The analytical specificity and sensitivity of the PCR primers were evaluated. The test showed ten-fold more sensitivity than conventional PCR and detected down to 275.5 fg/ µl of genomic DNA concentration, equivalent to 100 copies of KMT1 gene of P. multocida. The real-time PCR was found to be specific for KMT1 gene of P. multocida, as no cross reactivity was detected with a variety of known bacterial isolates. A total of 52 ovine lung tissue samples were screened for P. multocida, which showed improved level of detection as compared to conventional PCR. It is concluded that, this assay may be used as a valuable diagnostic tool for the rapid and specific detection of P. multocida. By virtue of its high throughput format and its ability to accurately identify as well as quantify the bacterial DNA, the method may be useful in large scale epidemiological studies and clarification of pathogenesis.
... In the evening, the animals returned back to the shed where they were provided with dry and green fodder and the formulated concentrates. Pneumonia was the leading cause of mortality at the farms [7,8]. Clinically, the sheep and goats exhibited coughing, sneezing, nasal discharge, and labored breathing. ...
The present study was planned to isolate Escherichia coli from pneumonic and septicemic sheep and goats and to determine
antimicrobial resistance (AMR) patterns of the isolates and evaluate them for the presence of extended-spectrum β-lactamase (ESBL)
using the phenotypic and molecular methods. Pneumonic lung tissues and heart blood samples were collected by performing necropsy
on sheep (n = 96) and goats (n = 08). The samples were processed and used for bacterial isolation. E. coli isolates (n = 58) including
53 from 34 sheep and 5 from 3 goats were recovered and identified by the cultural and biochemical characteristics and 16S rRNA
sequencing. The isolates were tested to ascertain AMR pattern, plasmid features, and ESBL production. The highest rate of resistance
(65.5%) was recorded against amoxicillin, enrofloxacin, norfloxacin, and ofloxacin. Eighteen (31%) out of 58 isolates showed the ESBL
phenotype by combination disk assay. There was a variability in the number (1–5) and size (2 to >20 kb) of plasmids among isolates.
Polymerase chain reaction (PCR) amplification revealed the presence of blaTEM (70.6%) and blaSHV (1.7%) genes. PCR specificity was
confirmed by nucleotide sequencing. The present study indicates that the extraintestinal and multidrug-resistant (MDR) E. coli strains
were associated with pneumonia and/or septicemia in sheep and goats. Therefore, thorough surveillance and monitoring of MDR
bacteria are urgently needed to implement the infection control strategies and to restore the efficacy of available antibiotics.
... Isolation and identification of M. haemolytica and P. multocida based on the cultural and biochemical characters and by the PCR assays targeting the specific genes have been reported earlier similar to the present study [13,31]. Moreover, the specific identification of P. multocida by 16S rRNA and KMT1 gene and the characterization by capsular typing has a significant impact on the epidemiology and control of pasteurellosis in domestic animals [8,32,33]. It is also proposed that the molecular typing methods provide rapid bacterial identification and have proved to be more specific over the cultural and biochemical tests [13,34]. ...
Aim: The purpose of this study was to isolate and characterize the Mannheimia haemolytica and Pasteurella multocida from blood, nasal discharge, and lung tissue of pneumonic goats.
Materials and Methods: A total of 14 goats were investigated for pneumonic pasteurellosis. Of 14 goats, nasal swabs and blood samples were collected from 10 clinically diseased animals. Moreover, lung tissue and heart blood samples were collected during necropsy of four goats died with pneumonia. All the samples were processed for the isolation of M. haemolytica and P. multocida in the laboratory. Bacterial isolates were identified by cultural and biochemical characters and 16S rRNA sequence analysis. All the isolates were subjected to susceptibility testing using commonly used antimicrobials. M. haemolytica isolates were characterized by PHSSA gene detection. P. multocida isolates were characterized by KMT1 gene detection and capsule typing.
Results: On necropsy of dead goats, the pneumonia was characterized as acute fibrinous bronchopneumonia. Bacterial culture revealed the isolation of M. haemolytica (7) and P. multocida (5) of 10 clinical cases. Moreover, M. haemolytica and P. multocida were coisolated from two of the lung tissues. Furthermore, one of the other two lung tissues showed the isolation of M. haemolytica while the other showed recovery of P. multocida. Bacterial isolates were specifically identified by the 16S rRNA sequence analysis. The isolates showed reduced susceptibility to β-lactams, aminoglycosides, and fluoroquinolones . Moreover, the PHSSA and KMT1 genes were specifically detected among M. haemolytica, and P. multocida isolates, respectively. All P. multocida isolates belonged to serogroup A.
Conclusion: The present study reported an occurrence of pneumonic pasteurellosis caused by M. haemolytica and P. multocida in a goat flock.
In this study, the presence and antimicrobial susceptibility of Mannheimia haemolytica and Pasteurella multocida pathogens in 200 sheep-lamb lung samples with pneumonia were investigated in 7 provinces of the Aegean region between 2019-2021. The tissues of the animals were inoculated into 7% blood agar and then incubated at 37oC for 24-48 hours. Gram staining was performed on the pure colonies and Gram-negative, oxidase-positive samples were confirmed with the Vitek 2 system. An Antibiogram test was performed and amoxicillin-clavulanic acid, enrofloxacin, erythromycin, florfenicol, gentamicin, oxytetracycline, sulfamethoxazole-trimethoprim, tulathromycin discs were used. M. haemolytica was detected in 20(10%) of the samples and P. multocida was detected in 22(11%) of the samples. 20 of the P. multocida isolates (91%) were susceptible to amoxicillin-clavulanic acid, florfenicol, and tulathromycin, and 20 of the M. haemolytica isolates (100%) were susceptible to enrofloxacin, oxytetracycline, florfenicol, and tulathromycin. Subsequently, 6 of the P. multocida isolates (27%) were resistant to erythromycin and oxytetracycline, and 5 of the M. haemolytica isolates (25%) were to sulfamethoxazole-trimethoprim. As a result, M. haemolytica and P. multocida can be seen with similar percentages as pneumonic agents in sheep-lamb pneumonia cases in the Aegean region, but effective treatment can be done with the right antibiotic selection.
The objective of this study was to evaluate the effects of crude glycerin (CG) on the diet of goats in feedlot, in terms of intake and nutrient digestibility, performance, feeding behavior, and metabolic profile. Forty castrated male goats with breed undefined and an average initial body weight of 19.7±2.3 kg were used. The experimental diets consisted of inclusion of CG at 0, 6, 12 and 18 % (based on DM). The DM (g/day), DM (g/kg), organic matter, crude protein, neutral detergent fiber, non-fiber carbohydrate, total digestible nutrients, and water intakes were decreased linearly with increasing CG levels, whereas ether extract intake increased linearly. The daily time spent (%) and average duration of events (h/d) for rumination presented a linear increase, although reduced for idling, as well as feeding and rumination efficiencies. The inclusion of CG did not affect the final body weight, total weight gain, and average daily gain. Serum cholesterol concentration presented a linear increasing effect. However, beta-hydroxybutyrate level was reduced. Crude glycerin containing 63.06% glycerol may partially replace corn and be included at a concentration of up to 18% of dry matter in the diet of finished goats in feedlot when moderate weight gains are desired, especially beneficial in semiarid regions.
