Bovine Host Research Articles

Hot topic: Influenza A H5N1 virus exhibits a broad host range, including dairy cows

The widespread circulation of highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b in wild birds in North America since late 2021 has resulted in multiple outbreaks in commercial and backyard poultry leading to major economic losses. Since the emergence of the virus in North America, multiple reassortment events have led to the emergence of many new variant genotypes that have been isolated from wild birds, with several viruses spilling over into poultry and other terrestrial and aquatic mammalian hosts. Notably, the most recent emerging HPAI H5N1 reassortant genotype B3.13 spilled over into dairy cattle (Bos taurus), resulting in unprecedented efficient transmission of the virus in this species, the first case of sustained transmission in a mammalian species. The transmission pathways involved in the spread of the virus from its first detection in Texas to several other states are complex. However, movement of subclinically infected cattle likely played a major role in virus spread. Infection in dairy cattle is characterized by the virus's tropism for milk-secreting cells in the mammary gland, leading to high viral load and shedding in milk. Replication of the virus in milk-secreting cells results in destruction of infected cells leading to severe viral mastitis, which is characterized by marked changes in milk quality (altered consistency and color) and pronounced decline in milk production by clinically affected animals. Here, we provide an overview of the HPAI H5N1 panzootic virus and discuss its host range and the current knowledge of its pathogenesis in the new bovine host.

Lipopolysaccharide Core Truncation in Invasive Escherichia coli O157:H7 ATCC 43895 Impairs Flagella and Curli Biosynthesis and Reduces Cell Invasion Ability.

Escherichia coli O157:H7 (E. coli O157) is known for causing severe foodborne illnesses such as hemorrhagic colitis and hemolytic uremic syndrome. Although E. coli O157 is typically regarded as an extracellular pathogen and a weak biofilm producer, some E. coli O157 strains, including a clinical strain ATCC 43895, exhibit a notable ability to invade bovine crypt cells and other epithelial cells, as well as to form robust biofilm. This invasive strain persists in the bovine host significantly longer than non-invasive strains. Various surface-associated factors, including lipopolysaccharides (LPS), flagella, and other adhesins, likely contribute to this enhanced invasiveness and biofilm formation. In this study, we constructed a series of LPS-core deletion mutations (waaI, waaG, waaF, and waaC) in E. coli O157 ATCC 43895, resulting in stepwise truncations of the LPS. This approach enabled us to investigate the effects on the biosynthesis of key surface factors, such as flagella and curli, and the ability of this invasive strain to invade host cells. We confirmed the LPS structure and found that all LPS-core mutants failed to form biofilms, highlighting the crucial role of core oligosaccharides in biofilm formation. Additionally, the LPS inner-core mutants ΔwaaF and ΔwaaC lost the ability to produce flagella and curli. Furthermore, these inner-core mutants exhibited a dramatic reduction in adherence to and invasion of epithelial cells (MAC-T), showing an approximately 100-fold decrease in cell invasion compared with the outer-core mutants (waaI and waaG) and the wild type. These findings underscore the critical role of LPS-core truncation in impairing flagella and curli biosynthesis, thereby reducing the invasion capability of E. coli O157 ATCC 43895.

Molecular identification and genetic variability of equine and bovine ocular setariasis in india: molecular profiling by mitochondrial genes.

Ocular setariasis is an ectopic infection caused by a parasite under the genus Setaria. Adult worms belong to the Setariidae family and typically reside in the peritoneal cavity of ungulates. However, immature forms of these species may aberrantly migrate to the eyes of cattle, buffalo, goats, horses and several other hosts, leading to corneal opacity and blindness. Here, we have distinguished the Setaria digitata collected from both equine and buffalo hosts based on the morphology, molecular profiling of mitochondrial cytochrome c oxidase subunit 1 (Cox1), cytochrome c oxidase subunit 3 (Cox3) and, Nicotinamide Adenine Dinucleotide dehydrogenase subunit 1 (NAD1) genes. A single filarial worm was collected from the eye of one equine and one bovine host. These worms were then processed for morphological examination and DNA isolation. Cox1, Cox3 and NAD1 genes were amplified using specific primers and subjected to custom sequencing. The sequences were then used for multiple sequence alignment, assessment of entropy, similarity and haplotype diversity analysis. Key morphological features confirmed the worms collected were male and female Setaria digitata from equine and buffalo hosts, respectively. Cox1, Cox3 and NAD1 gene sequence analysis showed a close association of S.digitata Indian isolates with its counterparts from Sri Lanka and China isolates. The phylogram of bovine S. digitata sequences shows a close relationship to other equine S. digitata sequences, indicating the need for further in-depth studies on the prevalence of infection across various host species and intermediate hosts. Although the sequence results suggest that S. digitata is likely the causative agent of ocular setariasis in India, additional samples are needed to confirm this conclusion. Comprehensive analysis of the transcriptome and proteome of S. digitata from both bovine and equine hosts is necessary to explore variations in host-parasite interactions. These findings will aid in future parasite identification, investigations into vector prevalence in India, and the development of control measures against ocular setariasis.

Draft genome sequences of 148 Klebsiella pneumoniae species complex members from bovine and human hosts.

Klebsiella pneumoniae species complex members, particularly K. pnemoniae sensu stricto, are common bovine clinical mastitis pathogens and often the cause of hospital- and community-acquired infections in humans. Here, we present 148 draft genome assemblies and annotations of K. pneumoniae species complex members from bovine and human hosts in Canada.

Expression network analysis of bovine skin infested with Rhipicephalus australis identifies pro-inflammatory genes contributing to tick susceptibility

The skin is the primary feeding site of ticks that infest livestock animals such as cattle. The highly specialised functions of skin at the molecular level may be a factor contributing to variation in susceptibility to tick infestation; but these remain to be well defined. The aim of this study was to investigate the bovine skin transcriptomic profiles of tick-naïve and tick-infested cattle and to uncover the gene expression networks that influence contrasting phenotypes of host resistance to ticks. RNA-Seq data was obtained from skin of Brangus cattle with high (n = 5) and low (n = 6) host resistance at 0 and 12 weeks following artificial tick challenge with Rhipicephalus australis larvae. No differentially expressed genes were detected pre-infestation between high and low resistance groups, but at 12-weeks there were 229 differentially expressed genes (DEGs; FDR < 0.05), of which 212 were the target of at least 1866 transcription factors (TFs) expressed in skin. Regulatory impact factor (RIF) analysis identified 158 significant TFs (P < 0.05) of which GRHL3, and DTX1 were also DEGs in the experiment. Gene term enrichment showed the significant TFs and DEGs were enriched in processes related to immune response and biological pathways related to host response to infectious diseases. Interferon Type 1-stimulated genes, including MX2, ISG15, MX1, OAS2 were upregulated in low host resistance steers after repeated tick challenge, suggesting dysregulated wound healing and chronic inflammatory skin processes contributing to host susceptibility to ticks. The present study provides an assessment of the bovine skin transcriptome before and after repeated tick challenge and shows that the up-regulation of pro-inflammatory genes is a prominent feature in the skin of tick-susceptible animals. In addition, the identification of transcription factors with high regulatory impact provides insights into the potentially meaningful gene–gene interactions involved in the variation of phenotypes of bovine host resistance to ticks.

Predicting hosts and cross-species transmission of Streptococcus agalactiae by interpretable machine learning

BackgroundStreptococcus agalactiae, commonly known as Group B Streptococcus (GBS), exhibits a broad host range, manifesting as both a beneficial commensal and an opportunistic pathogen across various species. In humans, it poses significant risks, causing neonatal sepsis and meningitis, along with severe infections in adults. Additionally, it impacts livestock by inducing mastitis in bovines and contributing to epidemic mortality in fish populations. Despite its wide host spectrum, the mechanisms enabling GBS to adapt to specific hosts remain inadequately elucidated. Therefore, the development of a rapid and accurate method differentiates GBS strains associated with particular animal hosts based on genome-wide information holds immense potential. Such a tool would not only bolster the identification and containment efforts during GBS outbreaks but also deepen our comprehension of the bacteria's host adaptations spanning humans, livestock, and other natural animal reservoirs. Methods and resultsHere, we developed three machine learning models—random forest (RF), logistic regression (LR), and support vector machine (SVM) based on genome-wide mutation data. These models enabled precise prediction of the host origin of GBS, accurately distinguishing between human, bovine, fish, and pig hosts. Moreover, we conducted an interpretable machine learning using SHapley Additive exPlanations (SHAP) and variant annotation to uncover the most influential genomic features and associated genes for each host. Additionally, by meticulously examining misclassified samples, we gained valuable insights into the dynamics of host transmission and the potential for zoonotic infections. ConclusionsOur study underscores the effectiveness of random forest (RF) and logistic regression (LR) models based on mutation data for accurately predicting GBS host origins. Additionally, we identify the key features associated with each GBS host, thereby enhancing our understanding of the bacteria's host-specific adaptations.

In vitro infection of bovine erythrocytes with Theileria annulata merozoites as a key step in completing the T. annulata life cycle in vitro

Theileria annulata is a protozoan parasite with a complex life cycle involving a bovine host and a tick vector. It is transmitted by Hyalomma ticks and is the causative agent of tropical theileriosis, a debilitating and often fatal disease in southern Europe, northern Africa and large parts of Asia. Understanding the biology of different life cycle stages is critical for the control of tropical theileriosis and requires the use of experimental animals which poses an ethical concern. We present for the first time the in vitro infection of red blood cells (RBCs) with T. annulata differentiated schizonts. The Ankara cell line of T. annulata was cultured at 41 °C for nine days to induce merogony and subsequently incubated with purified RBCs for one to three days. Percentage of parasitized erythrocyte (PPE) over the short culture period was estimated by Giemsa staining (0.007–0.01%), Flow cytometry activated sorting (FACS) (0.02–1.1%) and observation of FACS sorted cells by confocal microscopy (0.05–0.4%). There was a significant difference in the PPE between FACS and the two other techniques (one-way ANOVA followed by Tukey test, P = 0.004) but no significant difference was observed between the confocal imaging and Giemsa staining methods (ANOVA one-way followed by Tukey test, P = 0.06). Importantly, all three complementary methods confirmed the invasion of RBCs by T. annulata merozoites in vitro. Although the experimental conditions will require further optimization to increase the PPE, the in vitro infection of RBCs by T. annulata merozoites is pivotal in paving the way for the eventual completion of the T. annulata life cycle in vitro when combined with artificial tick feeding.

Comparison of the Automated OvaCyte Telenostic Faecal Analyser versus the McMaster and Mini-FLOTAC Techniques in the Estimation of Helminth Faecal Egg Counts in Equine.

Gastrointestinal helminth parasites continue to be a significant threat to the health of equine. OvaCyte Telenostic (OCT) (Telenostic Ltd., Kilkenny, Ireland) has developed an automated digital microscope utilising Artificial Intelligence to identify and count the clinically important helminth species in equine, bovine, and ovine host species. In this paper, the performance of the OCT analyser was evaluated for the detection and counting of equine helminth species parasites and its performance compared to the currently accepted benchmark methods of faecal egg counts being the McMaster and the Mini-FLOTAC techniques. A pairwise comparison of tests was assessed based on the correlation of egg counts and Cohen's kappa agreement statistics for dichotomized outcomes. Bayesian latent class analysis was used to estimate the sensitivity and specificity of all three techniques in the absence of a gold standard for four helminth parasites (strongyles, Anoplocephala spp., Parascaris spp. and Strongyloides westeri). Based on the analysis of 783 equine faecal samples, we found a high level (ρ ≥ 0.94) of correlation between each pairwise comparison of techniques for strongyle egg counts. Cohen's kappa agreement between techniques was high for strongyles and S. westeri, moderate for Parascaris spp., and low for Anoplocephala spp. All three techniques had a high sensitivity and specificity (>0.90) for strongyles. Across helminth parasites, the sensitivity of the OCT was the highest of the three techniques evaluated for strongyles (0.98 v 0.96 and 0.94), Anoplocephala spp. (0.86 v 0.44 and 0.46) and Parascaris spp. (0.96 v 0.83 and 0.96); but lowest for S. westeri (0.74 v 0.88 and 0.88), compared to McMaster and Mini-FLOTAC, respectively. In terms of specificity, OCT was the lowest in two species (Parascaris spp. 0.96, Anoplocephala spp. 0.95). In conclusion, OCT has a sensitivity and specificity statistically similar to both McMaster and Mini-FLOTAC, and had a higher correlation with Mini-FLOTAC. The OCT point of care faecal analyser offers improved workflow, test turn-around time and does not require trained laboratory personnel to operate or interpret the results.

Isolation and propagation of bovine blood-derived macrophages using a mixed culture with bovine endothelial B46 cells.

Macrophages are innate immune cells with multiple functions such as phagocytosis, cytokine production, and antigen presentation. Since macrophages play critical roles in some bacterial infectious diseases in cattle, including tuberculosis, paratuberculosis, and brucellosis, the in vitro culturing of bovine macrophages is useful for evaluating host-pathogen interactions at the cellular and molecular levels. We have previously reported the establishment of two immortalized bovine liver sinusoidal cell lines, endothelial B46 cells and myofibroblast-like A26 cells (Cell Biology International, 40, 1372-1379, 2016). In this study, we investigated the use of these cell lines as feeder cells that support the proliferation of bovine blood-derived macrophages (BBMs). Notably, the B46 cell line efficiently acts as feeder cells for the propagation of BBMs. Compared with primary cultured vascular endothelial cells, the infinite proliferation ability of B46 cells is more beneficial for preparing confluent feeder layers. In conclusion, this study provides a simple and efficient protocol for the isolation and propagation of BBMs using a primary mixed culture of bovine whole blood with B46 feeder cells. Isolated BBMs are expected to be useful for developing in vitro models for studying the interactions between bovine pathogens and host immune cells.

Differential modulation of innate immune response by lipopolysaccharide of Leptospira

Leptospirosis is a worldwide zoonosis caused by pathogenic Leptospira spp. having more than 300 serovars. These serovars can infect a variety of hosts, some being asymptomatic carriers and others showing varied symptoms of mild to severe infection. Since lipopolysaccharide (LPS) is the major antigen which defines serovar specificity, this different course of infection may be attributed to a differential innate response against this antigen. Previous studies have shown that Leptospira LPS is less endotoxic. However, it is unclear whether there is a difference in the ability of LPS isolated from different serovars to modulate the innate response. In this study, we purified LPS from three widely prevalent pathogenic serovars, i.e. Icterohaemorrhagiae strain RGA, Pomona, Hardjo, and from non-pathogenic L. biflexa serovar semeranga strain Potac 1 collectively termed as L-LPS and tested their ability to modulate innate response in macrophages from both resistant (mice) and susceptible (human and bovine) hosts. L-LPS induced differential response being more proinflammatory in mouse and less proinflammatory in human and bovine macrophages but overall less immunostimulatory than E. coli LPS (E-LPS). Irrespective of serovar, this response was TLR2-dependent in humans, whereas TLR4-dependent/CD14-independent in mouse using MyD88 adapter and signalling through P38 and ERK-dependent MAP kinase pathway. L-LPS-activated macrophages were able to phagocytose Leptospira and this effect was significantly higher or more pronounced when the macrophages were stimulated with L-LPS from the corresponding serovar. L-LPS activated both canonical and non-canonical inflammasome, producing IL-1β without inducing pyroptosis. Further, L-LPS induced both TNF-mediated early and NO-mediated late apoptosis. Altogether, these results indicate that L-LPS induces a differential innate response that is quite distinct from that induced by E-LPS and may be attributed to the structural differences and its atypical nature.

Sero-epidemiology of bovine tuberculosis in dairy cattle in Chattogram, Bangladesh

Mycobacterium bovis-caused bovine tuberculosis (bTB) is a global public health concern and commercially significant zoonotic disease. Many wild and domesticated animals, including cattle, are susceptible to this fatal disease and play an important role in cross-species transmission. To gain a better understanding of bTB in dairy cattle, a bTB serological study is also necessary in order to identify efficient bTB control strategies in Bangladesh. In Bangladesh, however, seroepidemiological research is either non-existent or limited (particularly Chattogram). To determine the seroprevalence of bTB and related possible risk factors in dairy cattle, a serological cross-sectional study was conducted (both animal and farm level). We estimated the seroprevalence of bTB was 7.5%(N=442) in this investigation, which also identified one animal-level and two farm-level possible risk factors. Individual animal risk factors included cattle source (own stock: OR=3.4, 95% CI: 1.4-8.1, p=0.006), and farm risk factors included population size (Larger: OR= 26.2, 95% CI: 2.2-319.1, p=0.010) and stocking density (Low and moderate: OR=19.6, 95%CI: 1.5-261.5, p=0.024). In the face-in housing system, there was also a higher risk of bTB seropositivity (OR=4.5). The research's key findings are thoroughly explored, and the findings will aid in the prevention and control of bTB in Bangladesh. In addition, early detection of positive cases, as well as their prompt removal and good farm management practices, can assist in preventing the spread of bTB to new human or bovine hosts. In underdeveloped nations like Bangladesh, a coordinated effort with local public health officials is critical for implementing an efficient TB control program.

The First Identification of Cryptosporidium parvum Virus-1 (CSpV1) in Hanwoo (Bos taurus coreanae) Calves in Korea.

Cryptosporidium is an obligate coccidian parasite that causes enteric diseases in bovine species. A double-stranded RNA virus associated with C. parvum oocysts, Cryptosporidium parvum virus-1 (CSpV1), has been characterized. However, the relationship between the abovementioned coccidian parasite and the virus has not been studied in the context of the known clinical outcomes. This study aimed to characterize the prevalence and molecular traits of CSpV1 in diarrheal feces of Hanwoo (Korean indigenous cattle) calves. Of the 140 fecal samples previously tested for C. parvum, which were obtained from Hanwoo calves aged 60 days, 70 tested positive and 70 tested negative. These samples were included in this study. By using the polymerase chain reaction (PCR) analysis targeting the RdRp gene of CSpV1, we detected CSpV1 in 28 samples (20.0%), with infection rates of 31.4% (22/70) in C. parvum-positive and 8.6% (6/70) in C. parvum-negative samples. CSpV1 samples detected in the same farm were clustered together. To the best of our knowledge, this is the first study to report the prevalence and molecular characteristics of CSpV1 in Hanwoo calves in the Republic of Korea, providing important insights into the relationship between C. parvum and CSpV1 in bovine hosts.

Genomic and phylogenetic profiling of RNA of tick-borne arboviruses in Hainan Island, China

Ticks act as vectors and hosts of numerous arboviruses. Examples of medically important arboviruses include the tick-borne encephalitis virus, Crimean Congo hemorrhagic fever, and severe fever with thrombocytopenia syndrome. Recently, some novel arboviruses have been identified in blood specimens of patients with unexplained fever and a history of tick bites in Inner Mongolia. Consequently, tick-borne viruses are a major focus of infectious disease research. However, the spectrum of tick-borne viruses in subtropical areas of China has yet to be sufficiently characterized. In this study, we collected 855 ticks from canine and bovine hosts in four locations in Hainan Province. The ticks were combined into 18 pools according to genus and location. Viral RNA-sequence libraries were subjected to transcriptome sequencing analysis. Molecular clues from metagenomic analyses were used to classify sequence reads into virus species, genera, or families. The diverse viral reads closely associated with mammals were assigned to 12 viral families and important tick-borne viruses, such as Jingmen, Beiji nairovirus, and Colorado tick fever. Our virome and phylogenetic analyses of the arbovirus strains provide basic data for preventing and controlling human infectious diseases caused by tick-borne viruses in the subtropical areas of China.

Staphylococcus microbes in the bovine skin microbiome attract blood-feeding stable flies

The human skin microbiome reportedly contributes to the attraction of mosquitoes to human hosts. We tested the hypothesis that bovine skin microbes affect the attraction of blood-feeding stable flies, Stomoxys calcitrans, to their bovine hosts. Microbes were collected from a calf and adult cow, and subsequently isolated and identified by mass spectrometry and genetic sequencing. Separate groups of (i) four Staphylococcus congeners (S. chromogenes, S. sciuri, S. simulans, S. succinus) and (ii) three bacterial heterogeners (Glutamicibacter protophormiae, Corynebacterium stationis, Wautersiella sp.) grown on agar, each attracted flies in still-air olfactometers, as did each Staphylococcus congener singly. The four Staphylococcus microbes also attracted flies in room bioassays. In greenhouse bioassays with paired black barrels as visual (surrogate host) stimuli, the treatment barrel baited with S. sciuri on agar induced significantly more fly alighting responses than the control barrel with sterile agar. This treatment effect could not be demonstrated on a cattle farm, possibly because of chemically and visually complex surroundings. Ammonia emitted by Staphylococcus microbes attracted flies, and a synthetic blend of microbe odorants enhanced the attractiveness of ammonia. Optimal attraction of stable flies to bovine microbes likely requires the integration of multimodal host cues.

Complete genomic characterization of bovine papillomavirus type 1 and 2 strains infers ongoing cross-species transmission between cattle and horses

Infection with bovine papillomavirus (BPV) types 1 and 2 results in the most common skin tumor of horses, termed equine sarcoid. The persistent and recurrent nature of this tumor stands in contrast to the regressive nature of BPV-1/− 2 induced cutaneous papillomas in cattle. The circulation of horse-specific BPV-1/− 2 variants within equine populations has been suggested as a possible explanation for the difference in clinical presentation of BPV-1/− 2 infection between horses and cattle. In order to investigate this hypothesis, we identified 98 complete BPV-1/− 2 genomes using a Nanopore sequencing approach. Separate BPV-1/− 2 alignments were used to infer Bayesian phylogenetic trees. Phylogeny-trait association concerning host species was investigated using Bayesian Tip-association Significance software (BaTS) Overall, 179 unique BPV-1 and 128 BPV-2 substitutions were found. The E2 coding region in the viral genome exhibited an exceptionally high rate of non-synonymous mutations (81 %, n = 13/16). Interestingly, extensive deletions in the L1/L2 region (up to 1.5 kb) were found exclusively in horse-derived samples. Nevertheless, the most frequently detected single nucleotide polymorphisms were shared between equine and bovine hosts, which is in agreement with BaTS results indicating no phylogeny-host correlation. We found indications that horse-specific mutations might exist in subpopulations of equine derived BPV-1/− 2, but these did not result in horse-adapted genetic variants. Based on these observations, cross-species transmission from cattle to horses seems to be an ongoing process, rather than an ancient occurrence that has been followed by the circulation of horse-adapted BPV variants in the horse population..

Blood meals from ‘dead-end’ vertebrate hosts enhance transmission potential of malaria-infected mosquitoes

Ingestion of an additional blood meal(s) by a hematophagic insect can accelerate development of several vector-borne parasites and pathogens. Most studies, however, offer blood from the same vertebrate host species as the original challenge (for e.g., human for primary and additional blood meals). Here, we show a second blood meal from bovine and canine hosts can also enhance sporozoite migration in Anopheles stephensi mosquitoes infected with the human- and rodent-restricted Plasmodium falciparum and P. berghei, respectively. The extrinsic incubation period (time to sporozoite appearance in salivary glands) showed more consistent reductions with blood from human and bovine donors than canine blood, although the latter's effect may be confounded by the toxicity, albeit non-specific, associated with the anticoagulant used to collect whole blood from donors. The complex patterns of enhancement highlight the limitations of a laboratory system but are nonetheless reminiscent of parasite host-specificity and mosquito adaptations, and the genetic predisposition of An. stephensi for bovine blood. We suggest that in natural settings, a blood meal from any vertebrate host could accentuate the risk of human infections by P. falciparum: targeting vectors that also feed on animals, via endectocides for instance, may reduce the number of malaria-infected mosquitoes and thus directly lower residual transmission. Since endectocides also benefit animal health, our results underscore the utility of the One Health framework, which postulates that human health and well-being is interconnected with that of animals. We posit this framework will be further validated if our observations also apply to other vector-borne diseases which together are responsible for some of the highest rates of morbidity and mortality in socio-economically disadvantaged populations.

Molecular typing of Staphylococcus aureus isolated from bovine and poultry based on PCR-RFLP of spa gene

Staphylococcus aureus (S. aureus) is considered one of the most important pathogen in bovine and poultry. The aim of the present study was molecular typing of S. aureus strains isolated from bovine mastitis and poultry by PCR-RFLP of spa gene. Fifty eight isolates of staphylococcus aureus were obtained from bovine mastitis (n=30) and poultry (n=28). Isolation identification were done based on conventional methods then confirmed by PCR method. The spa typing was performed by amplification of the spa gene X region. Bsp143I enzyme have been used for digestion of the PCR products and analyzed by electrophoresis on 1.5% gel. Gel Compare II software was used in pattern analysis and isolates with ≥ 70 percent similarities were taken as different types. Dendrogram obtained by RAPD results showed that among the 58 S. aureus strains eight RAPD types were identified (A–H). RAPD type A was more common among isolates from bovine mastitis (11 of 30 samples) and the majority (90%) of bovine isolates clustered in group A, D and E. Cluster B contains most S. aureus isolates of poultry origin. Group B, C and G clustered 82.1% of poultry host isolates, only in two clusters (F and H), some isolates from both the bovine and poultry host clustered together (7/58, 12%). In conclusion, RFLP of spa gene product using Bsp143I enzyme can be applied perfectly for differentiating and typing of S. aureus isolates were recovered from bovine mastitis and poultry infections.

Development of an enzyme linked immunosorbent assay using recombinant cathepsin B5 antigen for sero-surveillance of bovine tropical fasciolosis

Bovine tropical fasciolosis, caused by Fasciola gigantica, is a major parasitic disease in tropical countries responsible for significant production losses in animal husbandry practices. The disease is transmitted by the Radix sp. snails. In the early developmental stage of the parasite, the juveniles and immature flukes cause considerable damage to the liver parenchyma of the bovine host while migrating through the liver. The cathepsin (cat) B5 is a cysteine protease that is present in the excretory-secretory product of the fluke both in immature and adult stages. The early detection of fasciolosis is very critical in effective disease management. In this study, the cathepsin B5 gene from newly excysted juveniles were cloned, sequenced and analyzed. The phylogenetic analysis revealed existence of two distinct clades. The clade I includes cat B 1 to B3 whereas clade II consist of cat B4 to B7. Further, the present study was aimed to develop an enzyme linked immuno sorbent assay (ELISA) using recombinant cat B5 antigen. The developed enzyme immuno assay showed 95.3 % sensitivity and 92.4 % specificity with a cut-off of 60 % percent positive. It revealed weighted Kappa value as 0.768 (95 % CI 0.648–0.889) when compared with ELISA using native cathepsin protein. Hence, the developed assay can be exploited as a potent tool in the diagnosis and sero-surveillance of bovine tropical fasciolosis.

Repeated Emergence of Variant TetR Family Regulator, FarR, and Increased Resistance to Antimicrobial Unsaturated Fatty Acid among Clonal Complex 5 Methicillin-Resistant Staphylococcus aureus.

Resistance-nodulation-division (RND) superfamily efflux pumps promote antibiotic resistance in Gram-negative pathogens, but their role in Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) is undocumented. However, recent in vitro selections for resistance of S. aureus to an antimicrobial fatty acid, linoleic acid, and an antibiotic, rhodomyrtone, identified H121Y and C116R substitution variants, respectively, in a TetR family regulator, FarR, promoting increased expression of the RND pump FarE. Hypothesizing that in vivo selection pressures have also promoted the emergence of FarR variants, we searched available genome data and found that strains with FarRH121Y from human and bovine hosts have emerged sporadically in clonal complexes (CCs) CC1, CC30, CC8, CC22, and CC97, whereas multiple FarR variants have occurred within CC5 hospital-associated (HA)-MRSA. Of these, FarRE160G and FarRE93EE were exclusive to CC5, while FarRC116Y, FarRP165L, and FarRG166D also occurred in nonrelated CCs, primarily from bovine hosts. Within CC5, FarRC116Y and FarRG166D strains were polyphyletic, each exhibiting two emergence events. FarRC116Y and FarRE160G were individually sufficient to confer increased expression of FarE and enhanced resistance to linoleic acid (LA). Isolates with FarRE93EE were most closely related to S. aureus N315 MRSA and exhibited increased resistance independently of FarRE93EE. Accumulation of pseudogenes and additional polymorphisms in FarRE93EE strains contributed to a multiresistance phenotype which included fosfomycin and fusidic acid resistance in addition to increased linoleic acid resistance. These findings underscore the remarkable adaptive capacity of CC5 MRSA, which includes the polyphyletic USA100 lineage of HA-MRSA that is endemic in the Western hemisphere and known for the acquisition of multiple resistance phenotypes.

An exploration of within-herd dynamics of a transboundary livestock disease: A foot and mouth disease case study

Transboundary livestock diseases are a high priority for policy makers because of the serious economic burdens associated with infection. In order to make well informed preparedness and response plans, policy makers often utilize mathematical models to understand possible outcomes of different control strategies and outbreak scenarios. Many of these models focus on the transmission between herds and the overall trajectory of the outbreak. While the course of infection within herds has not been the focus of the majority of models, a thorough understanding of within-herd dynamics can provide valuable insight into a disease system by providing information on herd-level biological properties of the infection, which can be used to inform decision making in both endemic and outbreak settings and to inform larger between-herd models. In this study, we develop three stochastic simulation models to study within-herd foot and mouth disease dynamics and the implications of different empirical data-based assumptions about the timing of the onset of infectiousness and clinical signs. We also study the influence of herd size and the proportion of the herd that is initially infected on the outcome of the infection. We find that increasing herd size increases the duration of infectiousness and that the size of the herd plays a more significant role in determining this duration than the number of initially infected cattle in that herd. We also find that the assumptions made regarding the onset of infectiousness and clinical signs, which are based on contradictory empirical findings, can result in the predictions about when infection would be detectable differing by several days. Therefore, the disease progression used to characterize the course of infection in a single bovine host could have significant implications for determining when herds can be detected and subsequently controlled; the timing of which could influence the overall predicted trajectory of outbreaks.