Isolation and Identification of Pasteurella multocida and Mannheimia haemolytica from Pneumonic Small Ruminants and Their Antibiotic Susceptibility in Haramaya District, Eastern Ethiopia.

Background: Pasteurella species are frequently encountered as serious diseases in small ruminants. It is the main cause of respiratory pasteurellosis in sheep and goats of all age groups. Methods: The cross-sectional study was conducted from December 2022 to April 2023 in Haramaya district, eastern Ethiopia, to isolate and identify Pasteurella multocida and Mannheimia haemolytica and estimate their prevalence, associated risk factors, and antimicrobial sensitivity of isolates in small ruminants using a purposive sampling method. A total of 384 samples (156 nasal swabs from clinic cases and 228 lung swabs from abattoir cases) were collected. STATA 14 software was used to analyze the data. In addition, multivariable logistic regression analysis was performed to assess an association of risk factors. Results: Out of the 384 samples examined, 164 were positive for pasteurellosis, resulting in a 42.70% prevalence. Similarly, 63 (38.4%) of the 164 positive results were from nasal swabs, while 101 (61.6%) came from lung samples. M. haemolytica accounted for 126 (76.82%) of the isolates, while P. multocida accounted for 38 (23.17%). Of the 63 nasal swab isolates, 33 (37%) were from goats and 30 (42.8%) were from sheep. And 17 (10.89%) and 46 (29.58%), respectively, were P. multocida and M. haemolytica. Of the 46 (40%) of the 101 (44.3%) isolates of the pneumonic lung, samples were from goats, while 55 (48.47%) were from sheep. In this study, the risk factors (species, age, and body condition score) were found to be significant (p < 0.05). Pasteurella isolates evaluated for antibiotic susceptibility were highly resistant to oxacillin (90.90%), followed by gentamycin (72.72%), and penicillin (63.63%). However, the isolates were highly sensitive to chloramphenicol (90.90%), followed by tetracycline (63.63%), and ampicillin (54.54%). Conclusion: This study showed that M. haemolytica and P. multocida are the common causes of mannheimiosis and pasteurellosis in small ruminants, respectively, and isolates were resistant to commonly used antibiotics in the study area. Thus, an integrated vaccination strategy, antimicrobial resistance monitoring, and avoidance of stress-inducing factors are recommended.

Molecular and serological detection of Anaplasma spp. in small ruminants in an area of Cerrado Biome in northeastern Brazil.

Anaplasmosis, caused by bacteria of the genus Anaplasma, is an important tick-borne disease that causes economic losses to livestock farms in many countries. Even though Anaplasma spp. have been detected in goats and sheep worldwide, few studies investigate the occurrence and genetic identity of these agents in small ruminants from Brazil. Thus, this work aimed to detect and determine the genetic identity of Anaplasma spp. in small ruminants from the Baixo Parnaíba region, state of Maranhão, northeastern Brazil. For this purpose, blood samples were collected from 161 animals (91 goats; 70 sheep) from 4 municipalities in the Baixo Parnaíba region. Sheep and goat serum samples were subjected to recombinant membrane surface protein (MSP5)-based iELISA. Whole blood samples were subject to DNA extraction and molecular diagnosis using PCR assays for Anaplasma spp. targeting msp1ß, msp1α, 16S rRNA and msp4 genes. Positive samples were sequenced and then subjected to Anaplasma marginale msp1α genetic diversity analysis and phylogenetic inferences based on the 16S rRNA and msp4 genes. The serological survey detected the presence of anti-A. marginale IgG antibodies in 18 animals (11.1%): 2.9% (2/70) sheep and 17.4% (16/91) goats. Anaplasma marginale DNA was detected in 2 goats (1.2%) using qPCR based on the msp1ß gene. Two distinct A. marginale msp1α strains, namely α ß and α ß ΓγΓγΓγΓγ were found in the infected goats, each one found in a different animal, both belonging to the H genotype. Phylogenetic analysis based on the 16S rRNA gene showed the sequences positioned in three different clades and grouped with sequences from 'Candidatus Anaplasma boleense', A. platys and A. marginale. Phylogenetic inferences based on the msp4 gene positioned the sequence variants in the A. marginale clade. The present work represents the first molecular detection of sequence variants phylogenetic associated to 'Candidatus Anaplasma boleense' and A. platys and α ß and α ß ΓγΓγΓγΓγ in goats from Brazil.

Antibiotic treatment of Mycoplasma ovipneumoniae in domestic sheep (Ovis aries): Working at the livestock-wildlife interface in Yukon, Canada.

Domestic sheep (Ovis aries) can carry the bacterium Mycoplasma ovipneumoniae (M. ovipneumoniae) in their upper respiratory tract, often with little effect on health and productivity. However, for bighorn sheep (Ovis canadensis) populations, there is a link between M. ovipneumoniae infection and pneumonia, poor lamb recruitment, and high fatality rate. Because of these outcomes, preventing transmission of M. ovipneumoniae to free-ranging wild sheep has garnered interest from both the livestock and wildlife sectors. We hypothesized that treatment with intranasal and systemic enrofloxacin would reduce the prevalence of M. ovipneumoniae-positive animals in a flock of domestic sheep. Initially, the prevalence decreased in the treated group; but by 34 d post-treatment, the number of M. ovipneumoniae-positive sheep returned to near pretreatment prevalence. Key clinical message: Test-and-slaughter is a method used to reduce the risk of transmission of pneumonia-causing M. ovipneumoniae from domestic sheep and goats to free-ranging wild sheep. In an effort to find an alternative, we used enrofloxacin to treat a flock of M. ovipneumoniae-positive domestic sheep; however, long-term reduction of M. ovipneumoniae prevalence in the flock was not achieved.

Traitement antibiotique de Mycoplasma ovipneumoniae chez le mouton domestique (Ovis aries): travail à l'interface bétail-faune au Yukon, Canada. Les moutons domestiques (Ovis aries) peuvent être porteurs de la bactérie Mycoplasma ovipneumoniae (M. ovipneumoniae) dans leurs voies respiratoires supérieures, avec souvent peu d'effets sur la santé et la productivité. Cependant, pour les populations de mouflons d'Amérique (Ovis canadensis), il existe un lien entre l'infection à M. ovipneumoniae et la pneumonie, un faible recrutement d'agneaux et un taux de mortalité élevé. En raison de ces résultats, la prévention de la transmission de M. ovipneumoniae aux moutons sauvages en liberté a suscité l'intérêt des secteurs de l'élevage et de la faune sauvage. Nous avons émis l'hypothèse qu'un traitement par enrofloxacine intranasale et systémique réduirait la prévalence d'animaux positifs à M. ovipneumoniae dans un troupeau de moutons domestiques. Initialement, la prévalence a diminué dans le groupe traité; mais 34 jours après le traitement, le nombre de moutons positifs à M. ovipneumoniae est revenu à une prévalence proche de celle précédant le traitement.Message clinique clé :L'essai et l'abattage sont une méthode utilisée pour réduire le risque de transmission de M. ovipneumoniae, responsable de la pneumonie, des moutons et chèvres domestiques aux moutons sauvages en liberté. Dans le but de trouver une alternative, nous avons utilisé l'enrofloxacine pour traiter un troupeau de moutons domestiques positifs à M. ovipneumoniae; cependant, aucune réduction à long terme de la prévalence de M. ovipneumoniae dans le troupeau n'a été obtenue.(Traduit par Dr Serge Messier).

Exploring the microbial diversity and characterization of cellulase and hemicellulase genes in goat rumen: a metagenomic approach.

BACKGROUND: Goat rumen microbial communities are perceived as one of the most potential biochemical reservoirs of multi-functional enzymes, which are applicable to enhance wide array of bioprocesses such as the hydrolysis of cellulose and hemi-cellulose into fermentable sugar for biofuel and other value-added biochemical production. Even though, the limited understanding of rumen microbial genetic diversity and the absence of effective screening culture methods have impeded the full utilization of these potential enzymes. In this study, we applied culture independent metagenomics sequencing approach to isolate, and identify microbial communities in goat rumen, meanwhile, clone and functionally characterize novel cellulase and xylanase genes in goat rumen bacterial communities. RESULTS: Bacterial DNA samples were extracted from goat rumen fluid. Three genomic libraries were sequenced using Illumina HiSeq 2000 for paired-end 100-bp (PE100) and Illumina HiSeq 2500 for paired-end 125-bp (PE125). A total of 435gb raw reads were generated. Taxonomic analysis using Graphlan revealed that Fibrobacter, Prevotella, and Ruminococcus are the most abundant genera of bacteria in goat rumen. SPAdes assembly and prodigal annotation were performed. The contigs were also annotated using the DOE-JGI pipeline. In total, 117,502 CAZymes, comprising endoglucanases, exoglucanases, beta-glucosidases, xylosidases, and xylanases, were detected in all three samples. Two genes with predicted cellulolytic/xylanolytic activities were cloned and expressed in E. coli BL21(DE3). The endoglucanases and xylanase enzymatic activities of the recombinant proteins were confirmed using substrate plate assay and dinitrosalicylic acid (DNS) analysis. The 3D structures of endoglucanase A and endo-1,4-beta xylanase was predicted using the Swiss Model. Based on the 3D structure analysis, the two enzymes isolated from goat's rumen metagenome are unique with only 56-59% similarities to those homologous proteins in protein data bank (PDB) meanwhile, the structures of the enzymes also displayed greater stability, and higher catalytic activity. CONCLUSIONS: In summary, this study provided the database resources of bacterial metagenomes from goat's rumen fluid, including gene sequences with annotated functions and methods for gene isolation and over-expression of cellulolytic enzymes; and a wealth of genes in the metabolic pathways affecting food and nutrition of ruminant animals.

New insights into the genetic predisposition of brucellosis and its effect on the gut and vaginal microbiota in goats.

Goats contribute significantly to the global food security and industry. They constitute a main supplier of meat and milk for large proportions of people in Egypt and worldwide. Brucellosis is a zoonotic infectious disease that causes a significant economic loss in animal production. A case-control genome-wide association analysis (GWAS) was conducted using the infectious status of the animal as a phenotype. The does that showed abortion during the last third period of pregnancy and which were positive to both rose bengal plate and serum tube agglutination tests, were considered as cases. Otherwise, they were considered as controls. All animals were genotyped using the Illumina 65KSNP BeadChip. Additionally, the diversity and composition of vaginal and fecal microbiota in cases and controls were investigated using PCR-amplicone sequencing of the V4 region of 16S rDNA. After applying quality control criteria, 35,818 markers and 66 does were available for the GWAS test. The GWAS revealed a significantly associated SNP (P = 5.01 × 10-7) located on Caprine chromosome 15 at 29 megabases. Four other markers surpassed the proposed threshold (P = 2.5 × 10-5). Additionally, fourteen genomic regions accounted for more than 0.1% of the variance explained by all genome windows. Corresponding markers were located within or in close vicinity to several candidate genes, such as ARRB1, RELT, ATG16L2, IGSF21, UBR4, ULK1, DCN, MAPB1, NAIP, CD26, IFIH1, NDFIP2, DOK4, MAF, IL2RB, USP18, ARID5A, ZAP70, CNTN5, PIK3AP1, DNTT, BLNK, and NHLRC3. These genes play important roles in the regulation of immune responses to the infections through several biological pathways. Similar vaginal bacterial community was observed in both cases and controls while the fecal bacterial composition and diversity differed between the groups (P < 0.05). Faeces from the control does showed a higher relative abundance of the phylum Bacteroidota compared to cases (P < 0.05), while the latter showed more Firmicutes, Spirochaetota, Planctomycetota, and Proteobacteria. On the genus level, the control does exhibited higher abundances of Rikenellaceae RC9 gut group and Christensenellaceae R-7 group (P < 0.05), while the infected does revealed higher Bacteroides, Alistipes, and Prevotellaceae UCG-003 (P < 0.05). This information increases our understanding of the genetics of the susceptibility to Brucella in goats and may be useful in breeding programs and selection schemes that aim at controlling the disease in livestock.

Molecular surveillance and genetic diversity of Anaplasma spp. in cattle (Bos taurus) and goat (Capra aegagrus hircus) from Hainan island/province, China.

Anaplasmosis is a highly prevalent tick-borne intracellular bacterial disease that affects various host species globally, particularly ruminants in tropical and subtropical regions. However, information regarding the distribution and epidemiology of anaplasmosis in small and large ruminants on Hainan Isalnd is limited. To address this knowledge gap, the present study aimed to assess the occurrence of Anaplasma spp. infections in goats (N = 731) and cattle (N = 176) blood samples using nested PCR and conventional PCR based assays. The results revealed an overall prevalence of 30.1% in goats and 14.8% in cattle. The infection rates of A. bovis, A. phagocytophilum, A. ovis and A. capra in goat samples were 22.7%, 13.8%, 2.0% and 3.4%, respectively, while the infection rates of A. bovis, A. phagocytophilum and A. marginale in cattle samples were 11.4%, 6.3% and 5.7%, respectively. A. bovis exhibited the highest prevalence among the Anaplasma spp. in both goat and cattle samples. In addition, the most frequent co-infection was the one with A. phagocytophilum and A. bovis. It was found that the age, sex and feeding habits of cattle and goats were considered to be important risk factors. Evaluation of the risk factor relating to the rearing system showed that the infection rate for the free-range goats and cattle was significantly higher when compared with stall-feeding system.This study represents one of the largest investigations on the distribution, prevalence, and risk factors associated with Anaplasma infection in ruminants on Hainan Island, highlighting a higher circulation of the infection in the region than previously anticipated. Further reasesrch is necessary to investigate tick vectors, reservoir animals, and the zoonotic potential of the Anaplasma spp. in this endemic region of Hainan Island.

Epidemiology and Phylogeny of Anaplasma ovis with a Note on Hematological and Biochemical Changes in Asymptomatic Goats Enrolled from Four Districts in Punjab, Pakistan.

Background: Anaplasma ovis is an intra-erythrocytic gram negative rickettsial bacterium that infects small ruminants, resulting in huge economic losses worldwide. Materials and Methods: The present investigation aims at reporting the molecular prevalence of A. ovis in 1200 asymptomatic goats that were enrolled from 4 districts (Layyah, Lohdran, Dera Ghazi Khan, and Rajanpur) in Punjab, Pakistan by targeting the msp4 gene of bacterium. Risk factors associated with the prevalence of A. ovis and phylogeny of bacterium were also documented. Results: 184 out of 1200 (15%) goat blood samples were infected with A. ovis. The prevalence of the pathogen varied with the sampling sites (p = 0.005), and the highest prevalence was detected in goats from Layyah (19%) followed by Rajanpur (17%), Dera Ghazi Khan (15%), and Lohdran district (9%). The represented partial msp4 gene amplicon was confirmed by Sanger sequencing and deposited to GenBank (OP225957-59). Phylogenetic analysis revealed that the amplified isolates resembled the msp4 sequences reported from Iran, Mangolia, Sudan, and the United States. Sex and age of goats, herd composition and size, and the presence of ticks on goats and dogs associated with herds were the rick factors associated with the prevalence of A. ovis. Red blood cells, lymphocytes (%), neutrophils (%), hemoglobin, and hematocrit levels in blood and Aspartate amino transferase, urea, and creatinine levels in serum were disturbed in A. ovis infected goats when compared with uninfected animals. Conclusion: We are reporting the prevalence of A. ovis in Pakistani goats from four districts of Punjab and these data will help in developing the integrated control policies against this tick-borne pathogen that is infecting our goat breeds.

Differences in serum metabolome profile explain individual variation in growth performance of young goats.

High growth rates and body weight are important traits of young dairy goats that can shorten generation intervals, improve animal performance, and increase economic benefits. In the present study, ninety-nine, 6-month-old, female goats were fed with the same diet and kept under the same management condition. The ten goats with highest average daily gain (ADG, HADG, 135.27 ± 4.59 g/d) and ten goats with lowest ADG (LADG, 87.74 ± 3.13 g/d) were selected to identify the key serum metabolites associated with ADG, and to investigate the relationships of serum metabolome profiles with digestive tract microbiota. The results showed that a total of 125 serum metabolites were significantly different between HADG and LADG. Of these, 43 serum metabolites were significantly higher levels in HADG, including D-ornithine, l-glutamine, L-histidine, carnosine, LysoPC (16:1(9Z)/0:0), DCTP and hydroxylysine, while, 82 serum metabolites were significantly higher levels in LADG, including P-salicylic acid and deoxycholic acid 3-glucuronide. Pathway analysis indicated that these different metabolites were mainly involved in amino acid and lipid metabolism. Furthermore, Spearman's rank correlation analysis revealed that these differential serum metabolites were correlated with ADG and ADG-related bacteria. Notably, serum hydroxylysine and L-histidine could be used as biomarkers for distinguishing HADG and LADG goats, with an accuracy of >92.0%. SIGNIFICANCE: Our study confirms that individual microbiota and metabolic differences contribute to the variations of growth rate in young goats. Some serum metabolites may be useful in improving the growth performance of young goats, which provides directions for developing further nutritional regulation in the goat industry to achieve healthy feeding and efficiency enhancement.

Listeriosis in a goat herd.

Two 3-week-old goat kids from a herd of ~50 to 60 goats were examined by a veterinarian. The goats were in lateral recumbency with an inability to rise. Unilateral cranial nerve deficiencies included cervical rotation, nystagmus, ptosis, facial paralysis, and absence of palpebral reflex. One of the 2 kids had a fever. The kids died and necropsy examinations were performed. Histopathology findings were highly suggestive of Listeria monocytogenes infection, which was confirmed by bacterial culture. This case suggests that listeriosis should be included in the differential diagnosis for goats with neurological signs even if they are not fed silage or haylage and are kept in a clean barn.

Listériose dans un troupeau de chèvres. Deux chevreaux de 3 semaines d'un troupeau d'environ 50 à 60 chèvres ont été examinés par un vétérinaire. Les chèvres étaient en décubitus latéral avec une incapacité à se lever. Les déficiences unilatérales des nerfs crâniens comprenaient une rotation cervicale, du nystagmus, une ptose, une paralysie faciale et l'absence de réflexe palpébral. Un des 2 chevreaux avait de la fièvre. Les chevreaux sont morts et des nécropsies ont été effectués. Les résultats de l'histopathologie étaient très évocateurs d'une infection à Listeria monocytogenes, qui a été confirmée par culture bactérienne. Ce cas suggère que la listériose devrait être incluse dans le diagnostic différentiel pour les chèvres présentant des signes neurologiques même si elles ne sont pas nourries avec de l'ensilage ou de l'ensilage préfané et sont gardées dans une étable propre.(Traduit par Dr Serge Messier).

A serum NMR metabolomic analysis of the Corynebacterium pseudotuberculosis infection in goats.

Caseous lymphadenitis (CLA), an infectious disease caused by Corynebacterium pseudotuberculosis in small ruminants, is highly prevalent worldwide. Economic losses have already been associated with the disease, and little is known about the host-pathogen relationship associated with the disease. The present study aimed to perform a metabolomic study of the C. pseudotuberculosis infection in goats. Serum samples were collected from a herd of 173 goats. The animals were classified as controls (not infected), asymptomatic (seropositives but without detectable CLA clinical signs), and symptomatic (seropositive animals presenting CLA lesions), according to microbiological isolation and immunodiagnosis. The serum samples were analyzed using nuclear magnetic resonance (1H-NMR), nuclear Overhauser effect spectroscopy (NOESY), and Carr-Purcell-Meiboom-Gill (CPMG) sequences. The NMR data were analyzed using chemometrics, and principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) were performed to discover specific biomarkers responsible for discrimination between the groups. A high dissemination of the infection by C. pseudotuberculosis was observed, being 74.57% asymptomatic and 11.56% symptomatic. In the evaluation of 62 serum samples by NMR, the techniques were satisfactory in the discrimination of the groups, being also complementary and mutually confirming, demonstrating possible biomarkers for the infection by the bacterium. Twenty metabolites of interest were identified by NOESY and 29 by CPMG, such as tryptophan, polyunsaturated fatty acids, formic acid, NAD+, and 3-hydroxybutyrate, opening promising possibilities for the use of these results in new therapeutic, immunodiagnosis, and immunoprophylactic tools, as well as for studies of the immune response against C. pseudotuberculosis. KEY POINTS: • Sixty-two samples from healthy, CLA asymptomatic, and symptomatic goats were screened • Twenty metabolites of interest were identified by NOESY and 29 by CPMG • 1H-NMR NOESY and CPMG were complementary and mutually confirming.

Molecular detection and phylogenetic analysis of Anaplasma platys-like and Candidatus Anaplasma boleense strains from Argentina.

The present study aimed at the molecular detection of Anaplasma spp. in different samples obtained from cattle, goats and free-living Rhipicephalus microplus ticks from Argentina. DNA of members of the Anaplasmataceae family was detected by different PCR assays. The phylogenetic analyses of the obtained partial DNA sequences of the 16 S rDNA gene resulted in the identification of two different Anaplasma spp.: (I) Anaplasma platys-like bacteria (in blood sample from cattle and pools of R. microplus larvae and (II) Candidatus Anaplasma boleense (in blood samples from goats and one pool of R. microplus larvae of R. microplus). Candidatus A. boleense was found in two provinces that belong to different biogeographic regions, which leads to the conclusion that this bacterium may be widely distributed in Argentina. Interestingly, both Anaplasma spp. were found in the same R. microplus population in Chaco province, indicating that these two strains of Anaplasma are circulating in the same tick population. The results of this work represent the first report of the circulation of A. platys-like bacteria and Ca. A. boleense in domestic ruminants and free-living R. microplus ticks in Argentina. Further studies to determine the prevalence of infection, dispersion, clinical impact, transmission routes and cross-reactivity in serological tests of both Anaplasma species are needed.

A proof-of-concept study to investigate the efficacy of heat-inactivated autovaccines in Mycobacterium caprae experimentally challenged goats.

This study aimed to assess the efficacy of a heat-inactivated Mycobacterium caprae (HIMC) vaccine in goats experimentally challenged with the same strain of M. caprae. Twenty-one goats were divided into three groups of seven: vaccinated with heat-inactivated Mycobacterium bovis (HIMB), with HIMC and unvaccinated. At 7 weeks post-vaccination all animals were endobronchially challenged with M. caprae. Blood samples were collected for immunological assays and clinical signs were recorded throughout the experiment. All goats were euthanized at 9 weeks post-challenge. Gross pathological examination, analysis of lung pathology using computed tomography, and bacterial load quantification in pulmonary lymph nodes (LN) by qPCR were carried out. Only HIMC vaccinated goats showed a significant reduction of lung lesions volume and mycobacterial DNA load in LN compared to unvaccinated controls. Both vaccinated groups showed also a significant reduction of the other pathological parameters, an improved clinical outcome and a higher proportion of IFN-γ-producing central memory T cells after vaccination. The results indicated that homologous vaccination of goats with HIMC induced enhanced protection against M. caprae challenge by reducing lung pathology and bacterial load compared to the heterologous vaccine (HIMB). Further large-scale trials are necessary to assess the efficacy of autovaccines under field conditions.

P22 protein complex in the serodiagnosis of animal tuberculosis: Antigenic stability and cross-reactivity with Corynebacterium pseudotuberculosis infection.

The P22 ELISA was recently developed for the serodiagnosis of animal tuberculosis. Herein, the stability of the P22 antigen in different presentations and storage conditions, and the cross-reactivity with Corynebacterium pseudotuberculosis infection in small ruminants were evaluated. For the stability assay, serum samples from cows, sheep, goats, alpacas, badgers, and wild boar were used in the P22 ELISA. The cross-reactivity analysis used sera from sheep and goats with caseous lymphadenitis (CLA). Differences in the immune recognition of P22 were found when the antigen was stored at 40 °C, but without altering the negative or positive status of each sample. P22 ELISA presented 5.71 % cross-reactivity when CLA-positive sheep were evaluated, but no cross-reaction was observed among CLA-positive goat serum samples. This study showed that the P22 protein complex is stable under different formulations and temperatures, and that the assay presents a low cross-reactivity with CLA.

Generalized tuberculosis due to Mycobacterium caprae in a red fox phylogenetically related to livestock breakdowns.

BACKGROUND: Tuberculosis (TB) due to Mycobacterium caprae is endemic in goat herds and free-ranging wild boars in Spain, causing infections in other livestock or wild animals to a lesser extent. TB infection in foxes is infrequently reported and they are usually considered spillover hosts of TB. CASE PRESENTATION: A blind, depressed and severely emaciated red fox (Vulpes vulpes) was admitted to a rehabilitation center. After clinical examination it was humanely sacrificed. At necropsy, generalized TB lesions were observed that were subsequently confirmed by histopathology along with a co-infection with canine distemper virus. M. caprae was isolated from mycobacterial culture and spoligotype SB0415 was identified. Whole genome sequencing (WGS) of the isolated M. caprae was carried out and single nucleotide polymorphisms (SNP) were compared with other sequences of M. caprae isolated from livestock and wildlife of the same area throughout the last decade. CONCLUSIONS: This is the first reported case of TB due to M. caprae in a fox in the Iberian Peninsula. WGS and SNP analysis, together with spatial-temporal investigations, associated this case with recent M. caprae outbreaks in cattle and goat herds of the area. The results indicated transmission of M. caprae between livestock and the fox, suggesting that this species may occasionally play a role in the epidemiology of animal TB.

Molecular detection and phylogeny of Anaplasma spp. closely related to Anaplasma phagocytophilum in small ruminants from China.

The genus Anaplasma comprises eight bacterial species that are obligate intracellular pathogens that affect human and animal health. The zoonotic species A. phagocytophilum is the causative agent of tick-borne fever in ruminants, and of granulocytic anaplasmosis in horses, dogs, and humans. Recently, novel strains related to A. phagocytophilum (A. phagocytophilum-like 1/Japanese variant and A. phagocytophilum-like 2/Chinese variant) have been identified. The aim of this study was to reveal the prevalence and phylogeny of A. phagocytophilum and related stains in small ruminants and ticks in China based on sequences of the 16S rRNA combined restriction fragment length polymorphism (RFLP) and groEL genes. PCR-RFLP and phylogenetic analyses based on the 16S rRNA gene showed the presence of A. phagocytophilum-like 1 and 2 variants in sampled animals from China, with prevalence rates of 22.6% (303/1338) and 0.7% (10/1338), respectively. Only A. phagocytophilum-like 1 DNA was found in Haemaphysalis longicornis. The phylogeny based on the groEL gene showed inclusion of A. phagocytophilum-like 1 and some A. phagocytophilum-like 2 strains in two unique clades distinct from, but related to, Japanese and Chinese strains of related A. phagocytophilum, respectively. One noteworthy result was that the SSAP2f/SSAP2r primers detected Ehrlichia spp. strains. Moreover, the A. phagocytophilum-like 1 and 2 strains should be considered in the differential diagnosis of caprine and ovine anaplasmosis. Further investigations should be conducted to provide additional epidemiological information about A. phagocytophilum and A. phagocytophilum-like variants in animals and ticks.

Molecular survey for tick-borne pathogens and associated risk factors in sheep and goats in Kano Metropolis, Nigeria.

Tick-borne pathogens (TBPs) pose an increased health and productivity risk to livestock in sub-Saharan Africa. Information regarding TBPs infecting small ruminants in Kano metropolis is scarce. Therefore, we investigated the molecular epidemiology of tick-borne pathogens of economic importance from sheep and goats in Kano, Nigeria using Polymerase chain reaction (PCR). A total of 346 blood DNA samples were collected from small ruminants and analyzed for TBPs using PCR and sequencing. Risk of infection was determined for age, sex, breed and animal species. Our results indicate the absence of piroplasmids (Babesia/Theileria) and Rickettsia spp. infections. The overall prevalence for Anaplasma spp. was 9.25% (32/346) with a higher prevalence in goats 13.59% (25/184) compared with sheep 4.32% (7/162). With respect to age of animals, goats >4 years had the highest prevalence of 32.45% (11/37) which differs significantly (P = 0.0059) compared with other age categories. Cross breed goats had a prevalence of 15.63% (5/32) compared with Kano brown breed 14.08 (20/142). Sex significant difference (P = 0.029) was observed in the goats with females having the highest prevalence 20.89% (14/67) compared with males 9.40% (11/117). Furthermore, with regards to sheep, no significant difference (P > 0.05) was observed with respect to age and breed. Finally, no significant difference (P > 0.05) was observed with the prevalence of Anaplasma spp. due to Body condition score (BCS) in both sheep and goats. Conclusively, the occurrence of TBPs in small ruminants is low. Continuous efforts in tick control must be sustained to ensure high productive yield and reduced disease burden associated with TBPs of sheep and goats in Kano metropolis.

Molecular characterization and antimicrobial susceptibility of Corynebacterium pseudotuberculosis isolated from skin abscesses of native Korean goats (Capra hircus coreanae).

AIMS: This study aimed to investigate the molecular characterization and antimicrobial susceptibility of Corynebacterium pseudotuberculosis from skin abscesses of Korean native black goats (KNBG, Capra hircus coreanae) in South Korea. METHODS AND RESULTS: A total of 83 isolates were recovered from skin abscesses of KNBG. Of these isolates, 74 isolates were identified as C. pseudotuberculosis by phospholipase D (PLD) gene-based PCR assay. Each of the isolates possessed all 18 virulence genes (FagA, FagB, FagC, FagD, SigE, SpaC, SodC, PknG, NanH, OppA, OppB, OppC, OppD, OppF, CopC, NrdH and CpaE). The genetic diversity of C. pseudotuberculosis isolates was assessed by the phylogenetic analysis using the concatenated sequences (3073 bp) of five housekeeping genes (fusA, dnaK, infB, groeL1 and leuA) for investigating their genetic diversity. In the results, the isolates belonged to three groups: group 1 (67 isolates), group 2 (one isolate) and group 3 (six isolates) within biovar ovis. However, the groups exhibited low genetic diversity (0.20%-0.41%). In the antimicrobial susceptibility test, most isolates were susceptible to tetracycline, vancomycin, chloramphenicol, ciprofloxacin, erythromycin, enrofloxacin, cefoxitin, ampicillin, gentamycin, cephalothin and doxycycline, whereas they were not susceptible to cefotaxime, trimethoprim and streptomycin. CONCLUSION: This results suggest the involvement of relatively few clones of C. pseudotuberculosis in Korea. Further, present isolates can threaten public health due to potentially virulent strains with all 18 virulence genes and non-susceptible strains to clinically important antibiotics (CIA) and highly important antibiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first to investigate the genetic diversity and potential pathogenicity of C. pseudotuberculosis biovar ovis isolates from skin abscesses of KBNG in South Korea, and could provide useful information in controlling its infections.

Genome Engineering of the Fast-Growing Mycoplasma feriruminatoris toward a Live Vaccine Chassis.

Development of a new generation of vaccines is a key challenge for the control of infectious diseases affecting both humans and animals. Synthetic biology methods offer new ways to engineer bacterial chassis that can be used as vectors to present heterologous antigens and train the immune system against pathogens. Here, we describe the construction of a bacterial chassis based on the fast-growing Mycoplasma feriruminatoris, and the first steps toward its application as a live vaccine against contagious caprine pleuropneumonia (CCPP). To do so, the M. feriruminatoris genome was cloned in yeast, modified by iterative cycles of Cas9-mediated deletion of loci encoding virulence factors, and transplanted back in Mycoplasma capricolum subsp. capricolum recipient cells to produce the designed M. feriruminatoris chassis. Deleted genes encoded the glycerol transport and metabolism systems GtsABCD and GlpOKF and the Mycoplasma Ig binding protein-Mycoplasma Ig protease (MIB-MIP) immunoglobulin cleavage system. Phenotypic assays of the M. feriruminatoris chassis confirmed the corresponding loss of H2O2 production and IgG cleavage activities, while growth remained unaltered. The resulting mycoplasma chassis was further evaluated as a platform for the expression of heterologous surface proteins. A genome locus encoding an inactivated MIB-MIP system from the CCPP-causative agent Mycoplasma capricolum subsp. capripneumoniae was grafted in replacement of its homolog at the original locus in the chassis genome. Both heterologous proteins were detected in the resulting strain using proteomics, confirming their expression. This study demonstrates that advanced genome engineering methods are henceforth available for the fast-growing M. feriruminatoris, facilitating the development of novel vaccines, in particular against major mycoplasma diseases.

Enhancing Metabolic Efficiency through Optimizing Metabolizable Protein Profile in a Time Progressive Manner with Weaned Goats as a Model: Involvement of Gut Microbiota.

Feeding a growing global population and lowering environmental pollution are the two biggest challenges facing ruminant livestock. Considering the significance of nitrogen metabolism in these challenges, a dietary intervention regarding metabolizable protein profiles with different rumen-undegradable protein (RUP) ratios (high RUP [HRUP] versus low RUP [LRUP]) was conducted in young ruminants with weaned goats as a model. Fecal samples were collected longitudinally for nine consecutive weeks to dissect the timing and duration of intervention, as well as its mechanism of action involving the gut microbiota. Results showed that at least 6 weeks of intervention were needed to distinguish the beneficial effects of HRUP, and HRUP intervention improved the metabolic efficiency of goats as evidenced by enhanced growth performance and nutrient-apparent digestibility at week 6 and week 8 after weaning. Integrated analysis of bacterial diversity, metabolites, and inferred function indicated that HRUP intervention promoted Eubacterium abundance, several pathways related to bacterial chemotaxis pathway, ABC transporters, and butanoate metabolism and thereafter elicited a shift from acetate production toward butyrate and branched-chain amino acid (BCAA) production. Meanwhile, three distinct phases of microbial progression were noted irrespective of dietary treatments, including the enrichment of fiber-degrading Ruminococcus, the enhancement of microbial cell motility, and the shift of fermentation type as weaned goats aged. The current report provides novel insights into early-life diet-microbiota axis triggered by metabolic protein intervention and puts high emphasis on the time window and duration of dietary intervention in modulating lifelong performance of ruminants. IMPORTANCE Precise dietary intervention in early-life gastrointestinal microbiota has significant implications in the long-life productivity and health of young ruminants, as well as in lowering their environmental footprint. Here, using weaned goats as a model, we report that animals adapted to high rumen-undegradable protein diet in a dynamic manner by enriching fecal community that could effectively move toward and scavenge nutrients such as glucose and amino acids and, thereafter, elicit butyrate and BCAA production. Meanwhile, the three dynamic assembly trajectories in fecal microbiota highlight the importance of taking microbiota dynamics into account. Our findings systematically reported when, which, and how the fecal microbiome responded to metabolizable protein profile intervention in young ruminants and laid a foundation for improving the productivity and health of livestock due to the host-microbiota interplay.

Zoonotic bacteria in clinically healthy goats in petting zoo settings of zoological gardens in Germany.

Goats and other small ruminants are frequently used as contact animals in petting zoo settings of zoological gardens. However, they are capable to carry a broad spectrum of zoonotic pathogens without clinical signs. In this study, we analysed the presence of different zoonotic pathogens in 300 clinically healthy goats from 14 zoological gardens in Germany. Rectal and nasal swabs were investigated with a series of cultural and molecular techniques. In addition, vaginal swabs of the 230 female goats were investigated for the presence of Coxiella burnetii by real-time PCR. Antibodies against C. burnetii were tested in milk and serum by ELISA. Campylobacter spp. were found in 22.7%, Shiga-toxigenic Escherichia coli in 20.0% and Arcobacter spp. were found in 1.7% of the tested 300 goats after culture from rectal swabs and subsequent PCR. One sample contained an Escherichia fergusonii isolate with a blaCTX-M-1 -encoded extended-spectrum beta-lactamase phenotype. Neither Yersinia spp. nor Salmonella spp. were found. Nasal swabs of 20.7% of the goats yielded Staphylococcus aureus including one mecC-positive methicillin-resistant isolate. Neither Yersinia spp. nor Salmonella spp. were found, and none of the 230 vaginal swabs was positive for C. burnetii. Attempts to detect dermatophytes failed. In conclusion, a possible risk of transmission of zoonotic bacteria from goats in petting zoos to visitors should be considered. Appropriate information and facilities for hand washing and disinfection should be provided in all zoological gardens using goats as contact animals due to the regular presence of zoonotic bacteria in the collection.