The Distribution of Escherichia coli Serovars,Virulence Genes,Gene Association and Combinations and Virulence Genes Encoding Serotypes in Pathogenic E. coli Recovered from Diarrhoeic Calves,Sheep and Goat

Ruminants, especially cattle, have been implicated as a principal reservoir of one of the enterovirulent Escherichia coli pathotypes. The detection of the virulence genes in diarrhoeic calves and small ruminants has not been studied in Egypt. To determine the occurrence, serotypes and the virulence gene markers, stx1, stx2, hylA, Flic_h7, stb, F41, K99, sta, F17, LT‐I, LT‐II and eae, rectal swabs were taken from diarrhoeic calves, sheep and goats and subjected to bacterial culture and PCR. The E. coli prevalence rate in the diarrhoeic animals was 63.6% in calves, 27.3% in goat and 9.1% in sheep. The 102 E. coli strains isolated from the calves, goat and sheep were 100% haemolytic non‐verotoxic and fitted into the Eagg group. The isolates belonged to seven O serogroups (O25, O78, O86, O119, O158, O164 and O157). The eae gene was detected in six of the strains isolated from the calves. The 102 bovine, ovine and caprine E. coli strains isolated in this study were negative for stx1, stx2, F41, LT‐I and Flic_h7 genes. The highest gene combinations were found to occur in the form of 24/102 isolates (23.5%) that carried the F17 gene predominantly associated with eaeA, hylA, K99 and Stb genes in the calves, while the hylA, K99 and Sta were the only genes found to be in conjunction in both calves and goats (6/102; 5.9% each). Our data show that in Egypt, large and small ruminants could be a potential source of infection in humans.     

Diagnosis of Peste des Petits Ruminants in Wild and Domestic Animals in Xinjiang,China, 2013–2016

Peste des petits ruminants viruses (PPRVs) re‐emerged in China at the end of 2013 and then spread rapidly into 22 provinces through movement of live goats and sheep. In this study, 96 samples of domestic animals and 13 samples of wildlife were analysed for the presence of PPRV infection by ELISA or RT‐PCR. Of 96 samples from sheep and goats, 91 were PPRV positive, whereas all of the 13 samples from three wild species, Capra ibex (Capra ibex sibirica), argali (Ovis ammon) and Goitered gazelle (Gazella subgutturosa), were found to be positive. Five wildlife‐origin isolates from the above samples were identified as the lineage IV by a multiple alignment of the partial sequences in N gene.     

Ophthalmoscopic Characteristics in Sheep and Goats: Comparative Study

The ocular fundus was examined in 40 goat eyes and 40 sheep eyes by studying ophthalmoscopic characteristics and retinograms. Similarities and differing characteristics were described. In common: tapetal colour; peripheral yellowish area surrounding the Winslow stars; unpigmented areas in the non‐tapetal zone; a great amount of myelin in the optic disc; the Bergmeister's papilla and the holoangiotic retinal vascular pattern. Differences: big size of the Winslow stars in goats; myelinizated fibre over the non‐tapetal zone in sheep; shape, position and myelin distribution of the optic disc; and the presence of a ‘primary artery’ in goats.     

Genetic diversity and zoonotic potential of rotavirus A strains in the southern Andean highlands,Peru

Interspecies transmission is an important mechanism of evolution and contributes to rotavirus A (RVA) diversity. In order to evaluate the detection frequency, genetic diversity, epidemiological characteristics and zoonotic potential of RVA strains in faecal specimens from humans and animals cohabiting in the same environment in the department of Cusco, Peru, by molecular analysis, 265 faecal specimens were obtained from alpacas, llamas, sheep and shepherd children, and tested for RVA by RT‐PCR. Genotyping was performed by multiplex PCR and sequence analysis. Rotavirus A was detected in 20.3% of alpaca, 47.5% of llama, 100% of sheep and 33.3% of human samples. The most common genetic constellations were G3‐P[40]‐I8‐E3‐H6 in alpacas, G1/G3‐P[8]‐I1‐E1‐H1 in llamas, G1/G3/G35‐P[1]/P[8]‐I1‐E1‐H1 in sheep and G3‐P[40]‐I1/I8‐E3‐H1 in humans. The newly described genotypes P[40] and P[50] were identified in all host species, including humans. Genotyping showed that the majority of samples presented coinfection with two or more RVA strains. These data demonstrate the great genetic diversity of RVA in animals and humans in Cusco, Peru. Phylogenetic analysis suggested that the strains represent zoonotic transmission among the species studied. Due to the characteristics of the human and animal populations in this study (cohabitation of different host species in conditions of poor sanitation and hygiene), the occurrence of zoonoses is a real possibility.     

Evidence for Tunisian‐Like Pestiviruses Presence in Small Ruminants in Italy Since 2007

The genus Pestivirus, which belongs to the Flaviviridae family, includes ssRNA+ viruses responsible for infectious diseases in pigs, cattle, sheep, goats and other domestic and wild ruminants. Like most of the RNA viruses, pestivirus has high genome variability with practical consequences on disease epidemiology, diagnosis and control. In addition to the officially recognized species in the genus Pestivirus, such as BVDV‐1, BVDV‐2, BDV and CSFV, other pestiviruses have been detected. Furthermore, most of the ruminant pestiviruses show low or absent species specificity observed in serological tests and are able to infect multiple species. Particularly, small ruminants are receptive hosts of the most heterogeneous group of pestiviruses. The aim of this study was to carry out the molecular characterization of pestiviruses isolated from sheep and goats in Sicily, Italy. Phylogenetic analysis of two viral genomic regions (a fragment of 5′‐UTR and the whole N^pro regions) revealed the presence of different pestivirus genotypes in the analysed goat and sheep herds. Two of five viral isolates were clustered with BVDV‐1d viruses, a strain widespread in Italy, but never reported in Sicily. The other three isolates formed a distinct cluster with high similarity to Tunisian isolates, recently proposed as a new pestivirus species. This represents the first evidence for Tunisian‐like pestivirus presence in small ruminants in Italy. Furthermore, one of the isolates was collected from a goat, representing the first isolation of Tunisian‐like pestivirus from this species.     

Host range and geographical distribution of Babesia sp. Mymensingh

Bovine babesiosis represents a serious threat to the cattle industry in the tropics and subtropics. Although several Babesia species infect cattle, only B. bovis, B. bigemina and B. divergens are known to cause clinical babesiosis. However, our recent study demonstrated that the newly discovered Babesia sp. Mymensingh might be a virulent species capable of causing clinical babesiosis in cattle. The objective of this study was to determine the host range and geographical distribution of Babesia sp. Mymensingh on a global scale. A total of 2,860 archived DNA samples from 2,263 cattle in Sri Lanka (n = 672), the Philippines (n = 408), Vietnam (n = 460), Uganda (n = 409), Brazil (n = 164) and Argentina (n = 150); 419 buffalo in Sri Lanka (n = 327) and Vietnam (n = 92); and 127 goats and 51 sheep in Vietnam were screened using a Babesia sp. Mymensingh‐specific PCR assay. Babesia sp. Mymensingh infection was detected in cattle, buffalo, sheep and goats. Cattle of all countries surveyed in this study except Brazil were found to be infected with Babesia sp. Mymensingh. The highest positive rates were recorded in cattle from the Philippines (11.3%) and Vietnam (9.6%), followed by Argentina (4.7%), Sri Lanka (1.5%) and Uganda (1.0%). Buffalo were found to be infected with this parasite in Sri Lanka (1.2%) and Vietnam (10.9%). Unexpectedly, Babesia sp. Mymensingh was also detected in sheep (2.0%) and goats (1.3%) from Vietnam. These findings were confirmed by PCR amplicon sequencing. In conclusion, our present findings indicate that Babesia sp. Mymensingh, which infects cattle, buffalo, sheep and goats, is endemic in Asia, Africa and South America.     

Prevalence of Chlamydophila psittaci Infections in the Eyes of Cattle,Buffaloes, Sheep and Goats in Contact with a Human Population

This work is an example of cooperation between veterinary and human medicine being fully complementary and at the same time, indispensable to improve our knowledge on animal chlamydiosis. This study investigated the existence of ocular chlamydiae and determined the prevalence of its presence, chlamydiosis, in asymptomatic and diseased farm animals and adjacent humans. Data were obtained by the omp2 gene family Chlamydiaceae‐specific PCR. Two hundred cattle, buffaloes, sheep and goats and 44 human specimens were also examined. Conjunctival swabs from both the eyes were collected from all animals and humans using cotton swabs. Samples were tested for chlamydiae by Vero cells tissue culture, chicken embryo, modified Gimenez staining, direct fluorescein‐conjugated monoclonal antibody staining (FA), immunoperoxidase, CFT and PCR. The PCR‐RFLP revealed that Chlamydophila psittaci demonstrated in the conjunctival samples of cattle (68% asymptomatic and 88% diseased), of buffalo (68% asymptomatic and 72% diseased), of sheep (68% asymptomatic and 80% diseased), of goat (76% asymptomatic and 92% diseased) and of humans (77% asymptomatic and 82% diseased). The Cp. psittaci was the only chlamydiae demonstrated in all of the ocular conjunctival samples, which confirms the prevalence of Cp. psittaci in this population of animals and adjacent humans. Statistically, the animal species factor was calculated and was found to be of no significance. Yet, there appeared to be a significant difference in the percentage of animal that tested positive using the different methods. Detection of Cp. psittaci in most samples confirms the prevalence of Cp. psittaci in this population of animals and adjacent humans.     

Investigation of a Possible Link Between Vaccination and the 2010 Sheep Pox Epizootic in Morocco

Sheep pox is endemic in most parts of Northern Africa and has the potential to cause severe economic problems. Live attenuated vaccines are used in Morocco, and in many other countries, to control the disease. Sheep pox virus (SPPV) re‐appeared in 2010 causing a nodular clinical form previously not observed in Morocco. The severe clinical signs observed during the course of this outbreak and initial reports citing similarity in nucleotide sequence between the Moroccan vaccine strain and field isolates warranted a more in depth analysis of this epizootic. In this study, sequence analysis showed that isolates obtained from four provinces of eastern Morocco were identical, demonstrating that a single SPPV strain was responsible for the 2010 epizootic. In addition, the genome fragments sequenced and phylogenetic analyses undertaken as part of this study showed significant differences between field isolates and the Moroccan vaccine strain. New PCR methods were developed to differentiate between wild‐type isolates and vaccine strains of SPPV. Using these methods, no trace of wild‐type SPPV was found in the vaccine and no evidence was found to suggest that the vaccine strain was causing clinical disease.     

First detection and molecular identification of Sarcocystis spp. in small ruminants in North‐West Tunisia

Sarcocystosis is a parasitic disease caused by varying Sarcocystis species infecting humans and animals. It is commonly found in small ruminants causing pathogenic effects. This contributes to detrimental economic loss for local farmers and the local economy due this disease. Although the distribution of Sarcocystis can be found all over the world, the species infecting small ruminants in Tunisia is still unknown. Through this study, we aim to estimate the molecular prevalence of natural infection with Sarcocystis spp. in sheep and goats using molecular identification. Also, phylogenetic analyses were used to identify the different species of this parasite infecting small ruminants in northern Tunisia for the first time. DNA was extracted from 198 and 121, sheep and goats meat samples, respectively. The molecular prevalence of Sarcocystis spp. in sheep and goats was 58.6% (116/198) and 50.4% (61/121), respectively. Compared to the Noire de Thibar and cross‐breeds, the Barbarine sheep had the highest infection prevalence (63.4%) (p  =  .004). Five of the 116 positive samples were sequenced identifying Sarcocystis tenella from sheep. For goats, the sequencing results showed that five positive PCR products belonged to Sarcocystis capracanis species.     

Epidemiological investigation and phylogenetic analysis of caprine parainfluenza virus type 3 in sheep of China

Caprine parainfluenza virus type 3 (CPIV3) is a new member of the Respirovirus genus in the Paramyxivirudae family, mainly causing respiratory disease. Up to now, accumulating evidence has focused on CPIV3 infection in goats, with little understood about its epidemiology in sheep. To that end, we collected 1,163 sheep sera samples from nine provinces/autonomous regions in 2012 and 1,863 samples from six provinces/autonomous regions during 2016–2017, with serological prevalence of 50.3% (95% CI: 47.5, 53.3) and 64.9% (95% CI: 62.9, 67.2), respectively. Pathogenic detection by qRT‐PCR was also performed on serum samples collected in 2016–2017, and the percentage of CPIV3 positive samples was 21.5% (95% CI: 19.7, 23.5). Sequence alignment and phylogenetic analyses revealed 11 novel CPIV3 strains based on the M gene sequences. The M gene and full‐length sequences of CPIV3 strains derived from sheep shared high nucleotide similarity with goat‐origin strains, indicating conserved genome characteristics between the viruses. Furthermore, sequence evolution and epidemiological analysis show that CPIV3 is widespread throughout China. This is the first report describing CPIV3 infection in sheep in China, showing a high sero‐prevalence and contributes to the assessment of the epidemiology of CPIV3 in China.     

Serosurvey of Schmallenberg Virus Infections in Sheep and Goat Flocks in Lower Saxony,Germany

Schmallenberg virus (SBV) infections can cause congenital musculoskeletal and vertebral malformations as well as neurological failures in foetuses of several ruminant species if susceptible mother animals were infected during early gestation. Blood samples gained from 17 goat and 64 sheep flocks in Lower Saxony (LS), Germany (January–May 2012), which is located in the core region of the 2011/2012 epidemic were tested for antibodies against SBV by ELISA to detect past exposure to SBV. A SBV‐specific questionnaire was raised in all flocks. The calculated median within‐herd prevalence was 43.8% (min–max: 5.6–93.3%) for goats and 58.7% (min–max: 6.5–100%) for sheep, showing that small ruminants in LS, especially goats, are still at risk of novel SBV infections in the following lambing seasons as not all animals have seroconverted yet. Statistical analysis revealed that goats have a significantly lower risk of SBV infections than sheep which might be explained by different host preferences of Culicoides ssp. as main vectors for SBV and different housing conditions.     

Sequence and Phylogenetic Analyses of the Structural Genes of Virulent Isolates and Vaccine Strains of Peste Des Petits Ruminants Virus from India

Peste des petits ruminants (PPR) is an acute, highly contagious, notifiable and economically important transboundary viral disease of sheep and goats. In this study, sequence and phylogenetic analyses of structural protein genes, namely the nucleocapsid (N), the matrix (M), the fusion (F) and the haemagglutinin (H) coding sequences of virulent and vaccine strains of PPR virus (PPRV), were undertaken to determine the genetic variations between field isolates and vaccine strains. The open reading frame (ORF) of these genes of the isolates/strains was amplified by RT‐PCR, cloned and sequenced. The ORF of N, M, F and H genes was 1578, 1008, 1641 and 1830 nucleotides (nt) in length and encodes polypeptides of 525, 335, 546 and 609 amino acids (aa), respectively, as reported earlier. Comparative sequence analyses of these four genes of isolates/strains were carried out with published sequences. It revealed an identity of 97.7–100% and 97.7–99.8% among the Asian lineage IV and 89.6–98.7% and 89.8–98.9% with other lineages of PPRV at nt and aa levels, respectively. The phylogenetic analyses of these isolates based on the aa sequences showed that all the viruses belonged to lineage IV along with other Asian isolates. This is in agreement with earlier observations that only PPRV lineage IV is in circulation in India since the disease was first reported. Further, sequence analysis of the thermostable/thermo‐adapted vaccine strains showed no significant changes in the functional or structural surface protein–coding gene sequences. It is important to monitor the circulation of the PPRV in susceptible animals by H gene‐based sequence comparisons in addition to the F gene‐ and N gene‐based approaches to identify the distribution and spread of virus in the regular outbreaks that occur in endemic countries like India.     

Association among Results of Serum ELISA,Faecal Culture and Nested PCR on Milk,Blood and Faeces for the Detection of Paratuberculosis in Dairy Cows

Paratuberculosis is a chronic, infectious disease of ruminants that entails a serious concern for the cattle industry. One of the main issues relates to the efficiency of diagnosis of subclinically infected animals. The objective of this field study was to analyse the association among results of a serum enzyme‐linked immunosorbent assays (ELISA), faecal culture and nested PCR tests on milk, blood and faeces for Mycobacterium avium subsp. paratuberculosis detection in dairy cows. Faeces, blood and milk samples were collected from 328 lactating dairy cows in four known infected herds. Results were analysed to determine associations and levels of agreement between pairs of tests. A total of 61 animals (18.6%) tested positive when all the tests were interpreted in parallel. The agreement between results in different pairs of tests was poor, slight and fair in two, five and three of the 10 possible combinations respectively. Faecal culture and faecal polymerase chain reaction (PCR) resulted in the highest kappa coefficient (0.39; fair agreement), with the lowest agreement being for ELISA and blood PCR (−0.036; poor agreement). Fisher’s exact test resulted in statistically significant associations (P ≤ 0.05) between the following test pairs: ELISA : faecal culture; ELISA : faecal PCR; milk PCR : faecal PCR, blood PCR : faecal PCR and faecal culture : faecal PCR. Enzyme‐linked immunosorbent assays showed the highest complementary sensitivity values for all the possible two‐test combinations, followed by faecal PCR. The combined use of ELISA and faecal PCR has the potential to increase the overall sensitivity for the diagnosis of paratuberculosis infection.     

Clinical Presentation Resembling Mucosal Disease Associated with ‘HoBi’‐like Pestivirus in a Field Outbreak

The genus Pestivirus of the family Flaviviridae consists of four recognized species: Bovine viral diarrhoea virus 1 (BVDV‐1), Bovine viral diarrhoea virus 2 (BVDV‐2), Classical swine fever virus (CSFV) and Border disease virus (BDV). Recently, atypical pestiviruses (‘HoBi’‐like pestiviruses) were identified in batches of contaminated foetal calf serum and in naturally infected cattle with and without clinical symptoms. Here, we describe the first report of a mucosal disease‐like clinical presentation (MD) associated with a ‘HoBi’‐like pestivirus occurring in a cattle herd. The outbreak was investigated using immunohistochemistry, antibody detection, viral isolation and RT‐PCR. The sequence and phylogenetic analysis of 5′NCR, N^pro and E2 regions of the RT‐PCR positive samples showed that four different ‘HoBi’‐like strains were circulating in the herd. The main clinical signs and lesions were observed in the respiratory and digestive systems, but skin lesions and corneal opacity were also observed. MD characteristic lesions and a pestivirus with cytopathic biotype were detected in one calf. The present study is the first report of a MD like presentation associated with natural infection with ‘HoBi’‐like pestivirus. This report describes the clinical signs and provides a pathologic framework of an outbreak associated with at least two different ‘HoBi’‐like strains. Based on these observations, it appears that these atypical pestiviruses are most likely underdiagnosed in Brazilian cattle.     

Evaluation of effectiveness of Mass Vaccination Campaign against Peste des petits ruminants in Chhattisgarh state,India

The study evaluated the effectiveness of ‘Mass Vaccination Campaign (MVC)’ implemented against the contagious transboundary OIE notified Peste des petits ruminants (PPR) in sheep and goats on the lines of ‘pulse polio campaign’ for humans in Chhattisgarh state, India. The effectiveness was evaluated on the axes of adequacy, financial viability under with and without MVC through differencing under various scenarios and options and programme impact from a farmer's perspective. The adequacy evaluation revealed that the reported outbreaks, diagnosed and death cases declined under PPR‐MVC inconsonance with increased vaccination coverage. Furthermore, the seroconversion increased during post PPR‐MVC implies elevated immunity levels in the sheep and goat population. The estimated mean mortality loss was USD 45.2 and USD 16.5 per animal in goats and sheep, respectively, whereas the treatment and opportunity cost of labour was USD 1.9 and USD 2.5 per animal respectively. Under the low PPR incidence scenario, benefit: cost ratio, net present value and internal rate of return were 4.9:1, 48.9 million USD and 146.6%, whereas it was 12.4:1,142.7 million USD and 430.4% and 13.5:1,156.7 million USD and 430.4% under medium and high incidence scenarios. Furthermore, the option of vaccinating 100% risk population during the first year followed by 30% during subsequent years to cover naïve population will maximize benefits than 100% coverage every year; nevertheless, benefits outweighs cost manifolds in both of these options. The farmers had a positive opinion on the overall services provided under PPR‐MVC and the results provide the empirical evidence on effectiveness of ‘mass vaccination’ for its replication in other states of India or countries with similar socio‐economic and rearing environments.     

Pox outbreaks in Sheep and Goats at Makhdoom (Uttar Pradesh), India: Evidence of Sheeppox Virus Infection in Goats

Sheeppox and goatpox outbreaks occur often in India incurring huge economic loss to the small ruminant industry. This paper describes two sheeppox outbreaks, of which one occurred in an organized sheep breeding farm at Makhdoom (Uttar Pradesh), India, during 2007 and another in goats at the Central Institute of Research on Goats, Makhdoom (Uttar Pradesh), India during 2008. In the first outbreak, a local Muzaffarnagari sheep breed was affected (n = 477) with morbidity and mortality rates, respectively, of 100% and 53.9% accompanied by significant productivity losses. In the 2008 outbreaks, a small number of goats were affected without any mortality. The tissue and swabs collected from both the outbreaks were processed and inoculated onto Vero cells, and the causative agent of the outbreaks, capripox virus (CaPV), was isolated. The identity of the virus was confirmed as CaPV based on electron microscopy, experimental pathogenesis in sheep, capripox‐specific conventional and real‐time PCRs. Sequence analysis of the P32 envelope protein gene revealed that the causative agent of both outbreaks was confirmed as sheeppox virus (SPPV) implying SPPV infection not only in sheep but also goats in India.     

First Molecular Identification and Genetic Characterization of Theileria lestoquardi in Sheep of the Maghreb Region

Theileria lestoquardi is the most prominent Theileria species in small ruminants that causes malignant theileriosis of sheep in Africa and Asia. In the present survey, blood samples and ticks were collected in Kebili (southern Tunisia) from 166 Queue Fine de l'Ouest sheep. Giemsa‐stained blood smears, immunofluorescent antibody test (IFAT) and PCR were performed. The DNA was extracted from blood and analysed by PCR targeting 18S rRNA gene of Theileria spp. and then sequenced. A total number of 140 ticks were collected from a total number of 166 sheep during the four seasons. The ticks belonged to two genera and 4 species; the most frequent tick was Hyalomma excavatum 84.3% (118/140) and then Rhipicephalus spp. 15.7% (22/140). Only two animals had positive Giemsa‐stained blood smears, and they were also positive by IFAT. The amplicons had 99.3 and 99.6% homology with the BLAST published T. lestoquardi amplicons. To our knowledge, this is the first report of T. lestoquardi in small ruminants within the Maghreb region.