Ovis Infection Research Articles

Oestrose: uma parasitose emergente em pequenos ruminantes no Nordeste do Brasil

Resumo: No período de janeiro de 2011 a dezembro de 2014 foram diagnosticados 9 surtos (A, B, C, D, E, F, G, H e I) de Oestrus ovis em pequenos ruminantes no estado da Bahia. No surto A obteve-se 0,5% (1/200); B 2,2% (2/90); C 0,8% (1/120); D 2% (2/100); E 1% (1/100); F 3% (1/33); G 0,6% (1/150); H 2,5% (5/200); I com 11,4% (8/70) em ovinos e 5% (2/40) em caprinos. Os sinais clínicos associados ao parasitismo pelas larvas nos surtos foram respiração ruidosa, espirro seguido de secreção nasal catarral, inquietação, movimentação excessiva da cabeça e andar em círculo. Macroscopicamente havia nos seios e conchas nasais hiperemia, edema da mucosa e presença de larvas. Todas as larvas coletadas dos cornetos e conchas nasais variavam desde o primeiro ao terceiro estágio de desenvolvimento. Algumas larvas L3 coletadas nas necropsias foram incubadas e o imago obtido das pupas mediram aproximadamente 10mm de cor acinzentada e abdômen escurecido. Realizada análise descritiva das condições climáticas, ano e positividade de casos de oestrose, demonstrou que a ocorrência tem tendência de crescimento com os anos (p˂0,001) e que houve casos com menor média de temperatura mínima (p˂0,001), possibilitando o desenvolvimento da mosca de O. ovis, demonstrando que houve a introdução da mosca enTtre o rebanho de ovinos e caprinos do estado da Bahia, e que as condições climáticas são ideais para perpetuação da espécie.

Sperm quality in naturally infected rams with Brucella ovis

Ovine epididymitis caused by Brucella ovis is a chronic disease, characterized by testicular and epididymal alterations, which cause loss of fertility in rams. However, infected rams without clinical signs, may have decreased fertility and be excreting the bacteria. The aim of this study was to evaluate sperm quality in serologically positive to B. ovis naturally infected rams. Seminal samples were obtained of 207 rams to evaluate seminal volume, sperm concentration, mass motility, vitality, presence of somatic cells and percentage of abnormalities. Clinical examination was performed by palpation. The serological response against B. ovis was evaluated by double immunodiffusion in gel. The percentage variables were analyzed by GENMOD procedure and numerical variables were analyzed by ANOVA and differences were assessed using Tukey's test. 18.84% of the rams were positive against B. ovis. There was no difference in scrotal circumference and volume (P>0.05) between positive and negative animals for B. ovis. Sperm concentration, mass motility, vitality, and the percentage of sperm with abnormalities, were modified in positive animals (P<0.05), regardless of the presence of palpable lesions (P<0.05). The percentage of rams with lesions in the epididymis and severe presence of somatic cells was higher in positive animals (P<0.05). B. ovis infection adversely affects semen characteristics, due to an increase in epididymal lesions and by the severe presence of inflammatory cells in the ejaculate. Serological analysis to determine possible infection by B. ovis, should be considered as a component of the evaluation of fertility in rams, considering that sperm quality was affected in rams without lesions, but seropositive for B. ovis.

High infection of Anaplasma and Ehrlichia spp. among tick species collected from different geographical locations of Iran

ObjectiveTo ascertain the prevalence of the Anaplasma/Ehrlichia infections in tick population within four provinces of Iran. MethodsA total of 384 tick specimens were collected from domestic animals inhabiting in four provinces (East Azerbaijan, Gilan, South Khorasan and Yazd). Specimens were identified based on morphological analysis. The detection of Anaplasma spp./Ehrlichia spp. within tick samples was carried out by nested PCR amplification of the 16S ribosomal RNA gene accompanied by DNA sequencing and analysis for verification. ResultsA total of 10 tick species were identified as follows: Ornithodoros lahorensis (O. lahorensis) (44.8%), Hyalomma dromedarii (15.6%), Dermacentor marginatus (13.5%), Hyalomma anatolicum (11.2%), Hyalomma asiaticum (5.7%), Hyalomma marginatum (4.9%), Rhipicephalus sanguineus (2.3%), Hyalomma detritum (1.0%), Dermacentor niveus (0.5%) and Argas persicus (0.3%). The percentage distribution of Anaplasma/Ehrlichia was 55.5% (213) across 384 studied ticks. ConclusionsTo the best of our knowledge, this is the first report of Anaplasma ovis infection in O. lahorensis in Iran. We also conjecture the prevalence of Ehrlichia spp. in Yazd Province based on sequencing results; also, it is suggested that O. lahorensis is a potential vector in the studied area. This survey highlights the importance of Argasidae family to verify and correlate their threat in causing anaplasmosis and other diseases in animals.

The first outbreak of Taenia ovis infection in China

Infection of Taenia ovis metacestodes in sheep or goats causes great economic losses due to condemnation of carcasses. T. ovis infection is not formally recorded in China to date. In October, 2015, T. ovis infection occurred in Jingtai County, China, and 113 of 192 sheep from one farm were infected. Cysts resided in the cardiac and skeletal muscle, and evaginated metacestodes had four suckers and scolex armed with approximately 23 hooks. Using cox1 and nad1 as molecular markers, the sample was further identified and the results showed that the cox1 and nad1 nucleotide sequences of the sample shared 99% identity with that of T. ovis and 75%–91.3% with those of other Taenia species. Taken together, these results confirm the first occurrence of T. ovis in China.

EPERYTHROZOON OVIS INFECTIONS OF SHEEPAND GOATS IN SHALATEEN, ABU-RAMAD AND HALAEEB, RED SEA GOVERNORATE, EGYPT

An outbreak occurred in sheep and goats in triangular area (Shalateen –Abu-Ramad and Halaeeb), Red Sea Governorate, Egypt was described between January to December 2013.This outbreak showed clinical signs characterized by fever, in appetence, emaciation, anemia, icterus and ill-thrift. Clinical signs are most likely to be observed in young suckling and weaner animals. It is shown to be a complex problem attributable to many causes, some of which can be eliminated by supplementary feeding, drenching, vaccination, dipping and management. However, severe prolonged anemia accompanied by the development of ill-thrift still present in 248 animals (165 sheep and 83 goats). Infection with Eperythrozoon ovis was demonstrated in 21.42% and 12.50% of lambs and kids, 41.37 % and 28.75% of weaner sheep and goats and 10.75 % and 8.51% of adult sheep and goats respectively. When the general prevalence was analyzed by age, it was observed that weaners were mostly affected and disease was highly observed during winter and summer. Hematological examination showed macrocytic hemolytic anemia, with anisocytosis, poikilocytosis and marked left shift in erythrocyte maturation. Infected sheep and goats showed significant reduction in values of Hb, PCV and RBC while leukocytic count appeared slightly elevated. The main pathological changes in E. ovis infection were an increase in size of spleen at the peak of parasitaemia and an increase in liver size. Haemosiderin was present in livers, kidneys and spleens of all infected sheep and goats at the peak and late stages of parasitaemia.

Diversity of rickettsial pathogens in Columbian black-tailed deer and their associated keds (Diptera: Hippoboscidae) and ticks (Acari: Ixodidae)

Cervids host multiple species of ixodid ticks, other ectoparasites, and a variety of rickettsiae. However, diagnostic test cross-reactivity has precluded understanding the specific role of deer in rickettsial ecology. In our survey of 128 Columbian black-tailed deer (Odocoileus hemionus columbianus (Richardson)) and their arthropod parasites from two northern Californian herds, combined with reports from the literature, we identified four distinct Anaplasma spp. and one Ehrlichia species. Two keds, Lipoptena depressa (Say) and Neolipoptena ferrisi Bequaert, and two ixodid ticks, Ixodes pacificus Cooley and Kohls and Dermacentor occidentalis Marx, were removed from deer. One D. occidentalis was PCR-positive for E. chaffeensis; because it was also PCR-positive for Anaplasma sp., this is an Anaplasma/Ehrlichia co-infection prevalence of 4.3%. 29% of L. depressa, 23% of D. occidentalis, and 14% of deer were PCR-positive for Anaplasma spp. DNA sequencing confirmed A. bovis and A. ovis infections in D. occidentalis, A. odocoilei in deer and keds, and Anaplasma phagocytophilum strain WI-1 in keds and deer. This is the first report of Anaplasma spp. in a North America deer ked, and begs the question whether L. depressa may be a competent vector of Anaplasma spp. or merely acquire such bacteria while feeding on rickettsemic deer.

Evaluation of oxidant/antioxidant status, trace mineral levels, and erythrocyte osmotic fragility in goats naturally infected with Anaplasma ovis

Anaplasma ovis, an arthropod-borne pathogen that infects erythrocytes, is the major cause of ovine and caprine anaplasmosis. This study was performed to assess in goats infected with A. ovis the osmotic fragility of erythrocytes, antioxidant status, and serum levels of microminerals. Blood samples were collected from 104 mixed breed goats in Ahvaz area, southwest Iran and subjected to parasitologic, hematologic, oxidant/antioxidant, and micromineral assessment. Anaplasma infection was detected in 30 samples (28.8%) by microscopic examination of blood smears while PCR-RFLP analysis revealed infection with A. ovis in 68 samples (65.4%). Studied animals were divided into three groups based on A. ovis infection: Uninfected goats as control group (group 1), PCR positive without parasitemia (group 2) and PCR positive with parasitemia (group 3). Hematological evaluation showed significantly increased lymphocyte and monocyte counts in Anaplasma-infected groups (group 2 and 3). A significantly lower MCHC and higher MCV were also observed in infected groups. In group 3 significant rises in erythrocyte's osmotic fragility in different salt concentrations and also in median corpuscular fragility (MCF) was seen. Evaluation of the antioxidant defense system of the erythrocytes revealed a decrease in total antioxidant capacity (TAC) and superoxide dismutase (SOD) activity in group 3. There was no significant difference in serum micromineral levels between infected and uninfected animals. Overall, the observed substantial decrease in the antioxidant enzyme activities with remarkable elevated levels of erythrocyte osmotic fragility indicate high exposure of erythrocytes to oxidative damage in Anaplasma-infected goats. These results also suggest that the disturbed antioxidant defense mechanisms in caprine anaplasmosis can promote the development of anemia.

Serologic and molecular evidence of Brucella ovis infection in ovine and caprine flocks in the State of Minas Gerais, Brazil.

BackgroundBrucella ovis infection is one of the leading causes of sub fertility and infertility in ovine, been characterized mainly by epididymitis, orchitis and testicular atrophy in rams. This study aimed to determine the frequency of B. ovis positivity in rams and goats flocks in the State of Minas Gerais, Brazil, by agarose gel immunodiffusion (AGID), ELISA, Rose Bengal, PCR and bacteriological isolation as diagnostic tools.FindingsSerum and urine samples were collected from properties with sheep or goat flocks, or from properties with mixed flock. Out of 50 sheep flocks, 6 % (3/50) were seropositive by AGID while 4 % (2/50) were positive by urine PCR for B. ovis. Out of five goat farms, 20 % (1/5) were seropositive for B. ovis by AGID. Mixed flock farms had 11.1 % (2/18) of positivity by AGID. By ELISA, 19.5 % (8/41) of sheep properties and 61.1 % (11/18) of the properties with mixed flocks were positive for B. ovis. No samples were positive in the test of Rose Bengal, ruling out exposure to smooth LPS Brucella species (particularly Brucella melitensis) and indicating that the positive in the ELISA was associated with Brucella spp. LPS rough (presumably B. ovis). No urine sample from sheep or goat was positive by bacteriological isolation.ConclusionsOur results demonstrate serologic or molecular evidence of B. ovis infection in several rams and billy goats from meso-regions of the State of Minas Gerais, Brazil. Also, this study report the indirect ELISA as an important tool for the diagnosis of B. ovis infection, as indirect ELISA in this study demonstrated to be the most sensitive diagnostic method adopted.

Genetic variability, phylogenetic evaluation and first global report of Theileria luwenshuni, T. buffeli, and T. ovis in sheepdogs in Iran.

Theileriosis, caused by tick-borne hematozoan organisms, is a protozoan disease of domestic and wild mammals. The most phylo-molecular explorations of genus Theileria have been globally focused on the livestock and here are few investigations about canine Theileriosis. Twenty three and 52 blood samples were collected from sheepdogs and their owners, respectively, at different geographical foci of Mazandaran province (East, Central, and West regions), Northern Iran. To primary identify, the taken blood samples were fixed, stained, and surveyed by microscopic observation. DNA samples were extracted and amplified by polymerase chain reaction of 18s rRNA gene. The amplicons were directly sequenced to explore Theileria spp., and their heterogeneity traits. Sequencing and phylogenetic analyses revealed definitely the presence of Theileria luwenshuni, Theileria ovis, and Theileria buffeli (13%) in sheepdog isolates, while no infection was found in dog owners. Consensus positions of T. luwenshuni showed two amino acid substitutions (haplotype diversity; 0.9) in codons 53 and 64 compared to wild type with identity 99.5% and divergence 0.5-1.1, also one codon substitution (Hd; 0.875) found in T. buffeli with identity 97.6-99.5% and divergence 0.5-2.5. No novel haplotype was identified in T. ovis isolate. The current results strengthen our understanding about genetic variability, molecular taxonomic status, and epidemiology of canine theileriosis among sheepdogs and their owners in Iran where there is no similar study. As well, it will facilitate the implementation of measures to control theileriosis in the region. Based on our knowledge, the current study presents the first report of T. luwenshuni, T. buffeli, and T. ovis infections in sheepdogs worldwide.

Quantitative analysis of Babesia ovis infection in sheep and ticks

A quantitative PCR, based on the gene encoding Babesia ovis Surface Protein D (BoSPD) was developed and applied to investigate the presence of Babesia ovis (B. ovis) in its principal vector, the tick Rhipicephalus bursa (R. bursa), and in the ovine host. Quantification of B. ovis in experimentally-infected lambs showed a sharp increase in parasitemia 10–11days in blood-inoculated and adult tick-infested lambs, and 24days in a larvae-infested lamb. A gradual decrease of parasitemia was observed in the following months, with parasites detectable 6–12 months post-infection. Examination of the parasite load in adult R. bursa during the post-molting period using the quantitative PCR assay revealed a low parasite load during days 2–7 post-molting, followed by a sharp increase, until day 11, which corresponded to the completion of the pre-feeding period. The assay was then used to detect B. ovis in naturally-infected sheep and ticks. Examination of samples from 8 sheep and 2 goats from infected flocks detected B. ovis in both goats and in 7 out of the 8 sheep. Additionally, B. ovis was detected in 9 tick pools (5 ticks in each pool) and two individual ticks removed from sheep in infected flocks.

OCORRÊNCIA DE ANTICORPOS ANTI-Brucella ovis EM OVINOS DE SINOP E REGIÃO, MATO GROSSO, BRASIL

Neste estudo, foi realizado um levantamento soroepidemiológico da brucelose ovina por Brucella ovis em ovinos em idade reprodutiva da região norte-matogrossense com os objetivos de verificar frequência e distribuição da infecção por B. ovis em propriedades rurais e analisar os possíveis fatores de risco associados à infecção. Foram investigadas 11 propriedades criadoras de ovinos, das quais foram coletadas 337 amostras de sangue de carneiros e ovelhas de quatro municípios da mesorregião do Alto Teles Pires e Sinop, Mato Grosso. Os soros foram examinados pela técnica de imunodifusão em gel de ágar. De acordo com as análises, 72,7% (n=8) das propriedades apresentaram evidência sorológica de infecção por B. ovis, com uma frequência média de 13,9% (I.C.99%) de animais sororreagentes. Nas propriedades que higienizavam suas instalações com maior frequência e naquelas que realizavam criação no sistema extensivo, a soropositividade foi estatisticamente inferior (p&lt;0,05). Assim, neste estudo preliminar, verificou-se que a ovinocultura na região norte-matogrossense ainda não é uma atividade econômica primária, o que leva a uma baixa produtividade e gera predisposição à ocorrência da brucelose ovina e outras enfermidades em decorrência da baixa tecnificação da atividade.

Comparative immune responses against Psoroptes ovis in two cattle breeds with different susceptibility to mange.

The sheep scab mite, Psoroptes ovis, is a major problem in the beef cattle industry, especially in Belgian Blue (BB) cattle. This breed is naturally more predisposed to psoroptic mange but reasons for this high susceptibility remain unknown. Different immune responses could be a potential cause; thus in this study, the cutaneous immune response and in vitro cellular immune response after antigen re-stimulation were examined in naturally infested BB. Cytokine production in the skin and in circulating re-stimulated peripheral blood mononuclear cells (PBMC) demonstrated a mixed pro-inflammatory Th2/Th17 profile, with transcription of IL-4, IL-13, IL-6 and IL-17. Strong IL-17 up-regulation in the skin of BB was associated with an influx of eosinophils and other immune cells, potentially leading towards more severe symptoms. Virtually no changes in cutaneous IFN-γ transcription were detected, while there was substantial IFN-γ up-regulation in re-stimulated PBMC from infested and uninfested animals, potentially indicating a role of this pro-inflammatory cytokine in the innate immune response. In Holstein–Friesian (HF) cattle, generally more resistant to P. ovis infection, a largely similar immunologic response was observed. Differences between HF and BB were the lack of cutaneous IL-17 response in infested HF and low transcription levels of IFN-γ and high IL-10 transcription in re-stimulated PBMC from both infested and uninfested animals. Further research is needed to identify potential cell sources and biological functions for these cytokines and to fully unravel the basis of this different breed susceptibility to P. ovis.

Molecular Survey ofAnaplasmaSpecies in Small Ruminants Reveals the Presence of Novel Strains Closely Related toA. phagocytophilumin Tunisia

A survey of Anaplasma species in small ruminants is still lacking in North African countries. In this study, the presence of A. phagocytophilum, A. phagocytophilum-related species, and A. ovis was investigated in a total of 563 healthy small ruminants (303 goats and 260 sheep), from 25 randomly selected flocks sampled in Tunisia. Anaplasma spp. and A. ovis overall infection rates were 95.0% and 93.8% in sheep and 69.6% and 65.3% in goats, respectively. A. phagocytophilum was not detected in any of tested animals. A total of 20 sheep (7.7%) and 144 goats (47.5%) were infected by Anaplasma strains genetically related to A. phagocytophilum. Both in sheep and goats A. ovis prevalence was higher in adults (≥2 years) than in young (<2 years) subjects (p = 0.001 and 0.002 for goats and sheep, respectively). In sheep, A. ovis prevalence was higher in ewes with respect to rams (p = 0.010). The A. ovis infection rate was significantly lower in goats of the local breed (p = 0.049) and it was higher in goats infested by ticks than in not infested animals (p = 0.005). Genetic analysis of the msp4 gene of A. ovis indicated the presence of strains shared by Tunisian sheep and goats. Sequence analysis and phylogenetic studies on the basis of the 16S rRNA gene provided evidence for the circulation of at least two different potentially novel species genetically related to A. phagocytophilum in Tunisian small ruminants. These findings cause concern about specificity of serological tests used for detection of A. phagocytophilum in ruminants and provide additional information for elucidating pathogenesis and molecular epidemiology of A. phagocytophilum and related species.

The abcEDCBA-Encoded ABC Transporter and the virB Operon-Encoded Type IV Secretion System of Brucella ovis Are Critical for Intracellular Trafficking and Survival in Ovine Monocyte-Derived Macrophages

Brucella ovis infection is associated with epididymitis, orchitis and infertility in rams. Most of the information available on B. ovis and host cell interaction has been generated using murine macrophages or epithelial cell lines, but the interaction between B. ovis and primary ovine macrophages has not been studied. The aim of this study was to evaluate the role of the B. ovis abcEDCBA-encoded ABC transporter and the virB operon-encoded Type IV Secretion System (T4SS) during intracellular survival of B. ovis in ovine peripheral blood monocyte-derived macrophages. ΔabcBA and ΔvirB2 mutant strains were unable to survive in the intracellular environment when compared to the WT B. ovis at 48 hours post infection (hpi). In addition, these mutant strains cannot exclude the lysosomal marker LAMP1 from its vacuolar membrane, and their vacuoles do not acquire the endoplasmic reticulum marker calreticulin, which takes place in the WT B. ovis containing vacuole. Higher levels of nitric oxide production were observed in macrophages infected with WT B. ovis at 48 hpi when compared to macrophages infected with the ΔabcBA or ΔvirB2 mutant strains. Conversely, higher levels of reactive oxygen species were detected in macrophages infected with the ΔabcBA or ΔvirB2 mutant strains at 48 hpi when compared to macrophages infected with the WT strain. Our results demonstrate that B. ovis is able to persist and multiply in ovine macrophages, while ΔabcBA and ΔvirB2 mutations prevent intracellular multiplication, favor phagolysosome fusion, and impair maturation of the B. ovis vacuole towards an endoplasmic reticulum-derived compartment.

Encapsulated Brucella ovis Lacking a Putative ATP-Binding Cassette Transporter (ΔabcBA) Protects against Wild Type Brucella ovis in Rams.

This study aimed to evaluate protection induced by the vaccine candidate B. ovis ΔabcBA against experimental challenge with wild type B. ovis in rams. Rams were subcutaneously immunized with B. ovis ΔabcBA encapsulated with sterile alginate or with the non encapsulated vaccine strain. Serum, urine, and semen samples were collected during two months after immunization. The rams were then challenged with wild type B. ovis (ATCC25840), and the results were compared to non immunized and experimentally challenged rams. Immunization, particularly with encapsulated B. ovis ΔabcBA, prevented infection, secretion of wild type B. ovis in the semen and urine, shedding of neutrophils in the semen, and the development of clinical changes, gross and microscopic lesions induced by the wild type B. ovis reference strain. Collectively, our data indicates that the B. ovis ΔabcBA strain is an exceptionally good vaccine strain for preventing brucellosis caused by B. ovis infection in rams.

Oestrus ovis infection of grazing sheep during summer in southern Chile

Oestrus ovis is a botfly whose larvae cause nasal myiasis, an environmental-dependent disease in small ruminants, generating acute and chronic injuries in the cranial cavities of sheep. Chile is a country of the southernmost worldwide distribution of this parasite, and there is few information about. Whence, the objective of this study was to approximate the epidemiological situation of O. ovis infection in Chilean sheep. From December 2009 to March 2010, a total of 87 samples were obtained by necropsy for skull inspection. The larvae were collected, and microscopically identified. The prevalence in the sampled sheep was 60.9%. From those that were infected, 85.7% (18/21) of sheep were 1 to 3 years old, constituting the stratum with the highest prevalence. The difference of infection in females and males was not significant. The high risk of infection seems to be dependent upon the environmental conditions of this southern region, especially during summer when the first larval stage (L1) could be found as the evidence. Therefore, the disease should be considered as a significant problem for this kind of livestock production.

A study of hematological changes in sheep naturally infected with Anaplasma spp. and Theileria ovis: Molecular diagnosis

BACKGROUND: Ovine anaplasmosis and theileriosis are important tick-borne diseases of sheep and goats which are distributed in the tropical and subtropical areas of the world. OBJECTIVES: This study was performed to assess hematological status in sheep naturally infected with Anaplasma and Theileria spp. to clarify the pathogenic aspects of various species involved in ovine anaplasmosis and theileriosis in Ahvaz region. METHODS: 109 sheep were sampled, and blood parasite infections were diagnosed by microscopic examination and PCR. The blood samples were also subjected to hematologic assessment. RESULTS: PCR analysis revealed A. ovis infection in 86.2% of sheep, while mixed infections with A. marginale were also detected in 53.2% of them. However, Anaplasma inclusion bodies were only observed in 32.1% of the tested animals. T. ovis were found in 88% of the inspected sheep by PCR, and 67.8% of them were detected microscopically, as well. Hematologic assessment showed that mean RBC, PCV, Hb, and MCHC were significantly lower, whereas MCV and RDW were higher in the animals with mixed infections of Anaplasma with parasitemia and Theileria, compared to the uninfected sheep and groups with single infection or without parasitemia. CONCLUSIONS: In brief, it seems that Anaplasma can be activated and induce its pathogenesis in the presence of other infective agents in the carrier or asympthomatic animals. It can also be concluded that mixed infections of Anaplasma with parasitemia and Theileria may induce a regenerative anemia which is most likely attributable to a combined effect of the two.

Identification and expression of Babesia ovis secreted antigen 1 and evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay.

In order to identify immunoreactive proteins that are usable for the immunological diagnosis of Babesia ovis infections, a phage lambda cDNA expression library was constructed and screened using parasite-specific immune serum. Immunoscreening resulted in the identification of a full-length cDNA clone encoding a secreted protein designated Babesia ovis secreted antigen 1 (BoSA1). The full-length BoSA1 cDNA contained a 1,137-bp open reading frame that encoded a protein of 378 amino acids, with a signal peptide and 2 internal repeat domains. The theoretical molecular mass of the mature protein was 42.5 kDa. Recombinant BoSA1 (rBoSA1) protein was expressed in Escherichia coli strain DH5α cells as a glutathione S-transferase (GST) fusion protein and was purified by affinity chromatography. Purified rBoSA1 was tested for reactivity with sera from animals experimentally or naturally infected with B. ovis, in an indirect enzyme-linked immunosorbent assay (ELISA). The results showed that specific antibodies against rBoSA1 were detectable on days 7 and 8 of the experimental infection and were maintained during the sampling period. Additionally, 38 field sera taken from sheep naturally infected with B. ovis gave strong positive reactions in the ELISA between day 20 and day 30 of treatment. As a result, the identified recombinant BoSA1 protein seems to be a promising diagnostic antigen that is usable for the development of serological assays for the diagnosis of ovine babesiosis. This is the first report on the molecular cloning, expression, and potential use of a recombinant antigen for the diagnosis of ovine babesiosis.

Hemotropic Mycoplasma ovis infection in goats with concurrent gastrointestinal parasitism in Malaysia

Hemotropic Mycoplasmosis is common in sheep and goats worldwide, which leads to huge economic losses. In this study, ten goats each were sampled from five herds belonging to the Ladang Angkat, Faculty of Veterinary Medicine (FVM) for the assessment of Mycoplasma ovis infection and concomitant intestinal parasites burden. Giemsa stain and Modified McMaster techniques were used to study the hemotropic mycoplasmosis and gastrointestinal parasite burden, respectively. Questionnaires were equally administered to each farmer and a fly trap was used to trap biting flies around the goat herds. Out of 50 samples analyzed, 94.0% (n=47/50) were positive for M. ovis infection. Among the positive samples, 93.6% (n=44/47) were mild infection while 6.4% (n=3/47) were moderate infection, with highest infection rate of 38.5% parasitemia. There was a significant association (P

Microarray Analysis of Gene Expression in Rams Experimentally-Infected with the Virulent Strain of Brucella ovis

Microarray analysis of gene expression profiles of total mRNA was studied in rams experimentally-infected with the virulent strain of Brucella ovis (B. ovis). The tissues studied included reproductive organs (epididymus, testicles, ampolae, vesicular glands, bulbourethral glands) and a pool of lymph nodes (inguinal and scrotal). The microarray analysis of each tissue was done at three time points: acute infection (60 days post infection [dpi]), chronic infection phase I (120 dpi), and chronic infection phase II (240 dpi). The gene expression profiles associated to B. ovis infection were determined using the Affymetrix Bovine Genome Array and expression levels of infected and uninfected rams (control group, 0 dpi) were compared. Of the 23,000 genes analyzed in the microarrays, 139 during acute phase of infection, 930 during the chronic phase I and 744 genes from the chronic phase II were identified as Differentially Expressed Genes (DEGs). Thirty known genes and fourteen unknown genes were expressed during the three phases of infection in all tissues. The biological functions of these genes included immune cell trafficking, immunological disease, infectious disease, inflammatory disease, inflammatory response and cellular movement, and significant differences were observed at the three phases of infection and in all infected tissues. For the first analysis, 8 genes in common during the three time points and to all infected tissues were chosen for the microarray validation. Acute phase of infection supported the relevance of the ataxia telangiectasia-mutated ATMIN (ATM interactor) gene. RCHY1 (ring finger and CHY zinc finger domain containing 1), ANKFY1 (ankyrin repeat and FYVE domain containing 1) and EFEMP1 (EGF-containing fibulin-like extracellular matrix protein 1) were also highlighted by the analysis during chronic phase I of infection. ZRAB2 (Zinc finger, RAN-binding domain containing 2) was validated at chronic phase II. This study represents the first analysis of microarray gene expression in different tissues of rams infected with of B. ovis at various times during infection and many of the genes identified require further study. These results expand the knowledge of the pathogenesis of this B. ovis strain infection in rams and suggest new genes and pathways for further investigation.