Bovine Abortion Research Articles

Conventional and molecular diagnosis of Campylobacteriosis associated with bovine abortion

Campylobacteriosis is responsible for genital tract infections of beef and dairy herds, causing a significant economic loss in livestock sector. Campylobacter foetus species is one of the important pathogens because of its potential impact in Veterinary and Human health. This study was designed to determine the regional incidence of C. foetus infection in Kerala, India by isolation, detection of C. foetus in clinical samples by Polymerase Chain Reaction (PCR), real time PCR (qPCR), and Enzyme Linked Immunosorbent Assay (ELISA) for the detection of C. foetus antibodies in sera of bovines with the history of abortion/infertility. Clinical samples (aborted materials (50), serum (50), Cervico-Vaginal Mucus (CVM) (30) and semen samples (30)) from a total of 160 cattle and buffaloes with the history of abortion and infertility were collected. Aborted materials including placenta, foetal membranes, liver, lungs and stomach contents of the aborted foetus, semen and CVM samples were processed and subjected to isolation and identification of Campylobacter foetus subsp. foetus (Cff) and Campylobacter foetus subsp. venerealis (Cfv) and molecular confirmation by PCR and qPCR respectively. Serum samples from aborted dams were tested using indirect ELISA. All the suspected clinical samples were found negative for Cff and Cfv on both culturing and PCR. All the serum samples tested were negative by ELISA as well. Conclusively the study indicated the infection of C. foetus spp. responsible for abortion in bovine are rare in the location where the study was conducted, which might be due to insignificant endemic levels. As per the breeding policy, only artificial insemination is practiced in Kerala in bovines, which is often considered as a simple control method for Bovine Genital Campylobacteriosis (BGC) and might be one of the factors that prevented extensive spread of C. foetus spp. infection.

Retrospective study of factors associated with bovine infectious abortion and perinatal mortality

Abortion and perinatal mortality, leading causes of economic loss in cattle industry, are the consequence of both non-infectious and a wide range of infectious causes. However, the relative contribution of pathogens to bovine abortion and perinatal mortality is poorly documented, since available studies involved only a limited number of pathogens. Therefore, the objectives of the present monitoring study were to determine the prevalence of infectious agents associated with bovine abortion and perinatal mortality, and to identify differences in production type, gestation length, parity and seasonality by using mixed effect models (logistic regression). A pre-established sampling protocol based on the collection of the aborted fetus/calf and a corresponding maternal blood sample, involving diagnostic testing for 10 pathogens, was performed. At least one potential causal agent of the abortion or perinatal mortality was detected in 39 % of cases. In these diagnosed cases, Neospora caninum was the most detected pathogen, followed by Trueperella pyogenes, BVDv, Escherichia coli, and Aspergillus fumigatus. Neospora caninum [odds ratio (OR): 0.4; 95 % confidence interval (CI): 0.3–0.7] and Aspergillus fumigatus (OR: 0.1; 95 % CI: 0.1–0.3) were detected less in late versus early gestation. Aspergillus fumigatus was less common in dairy in comparison to beef abortion cases (OR: 0.2; 95 % CI: 0.1–0.6). Winter was associated with a lower positivity for Neospora caninum and BVDv in comparison to warmer seasons. Despite extensive diagnostic testing, an etiological diagnosis was not reached in 61 % of cases, highlighting the need for even more extensive (non-)infectious disease testing or more accurate tests.

Genetic characterization of Neospora caninum from aborted bovine fetuses in Hokkaido, Japan.

Neospora caninum is one of the main causes of bovine abortions worldwide, including Japan. Nothing is known about the N. caninum population substructures in Japan, and only one isolate from a pregnant sheep has been studied to date. This study describes, for the first time, the genetic characterization of isolates of N. caninum implicated in cattle abortions in Japan. Brains from five aborted fetuses were successfully genotyped based on multilocus microsatellite markers. Assigned genotypes showed high frequencies of mixed alleles in the sequenced markers MS7 and MS10, raising concerns about the subpopulation structures of N. caninum infecting animals in Japan. Clustering analysis of the genotypes, together with those from a previous dataset, showed that five of the six genotypes were distinct from other clusters. Meanwhile, the remaining genotype, together with the sheep isolate from Japan, was grouped with those from Mexico and Spain. These preliminary data may indicate a complex transmission pattern of N. caninum in Japan via clonal spreading by vertical and horizontal transmission and geographically related population substructuring.

Bovine Abortions Revisited-Enhancing Abortion Diagnostics by 16S rDNA Amplicon Sequencing and Fluorescence in situ Hybridization.

Abortion in cattle causes significant economic losses for cattle farmers worldwide. The diversity of abortifacients makes abortion diagnostics a complex and challenging discipline that additionally is restrained by time and economy. Microbial culture has traditionally been an important method for the identification of bacterial and mycotic abortifacients. However, it comes with the inherent bias of favoring the easy-to-culture species, e.g., those that do not require cell culture, pre-enrichment, a variety of selective growth media, or different oxygen levels for in vitro growth. Molecular methods such as polymerase chain reaction (PCR) and next-generation sequencing have been established as alternatives to traditional microbial culturing methods in several diagnostic fields including abortion diagnostics. Fluorescence in situ hybridization (FISH), a bridging microscopy technique that combines molecular accuracy with culture independence, and spatial resolution of the pathogen-lesion relation, is also gaining influence in several diagnostic fields. In this study, real-time quantitative PCR (qPCR), 16S rDNA amplicon sequencing, and FISH were applied separately and in combination in order to (i) identify potentially abortifacient bacteria without the bias of culturability, (ii) increase the diagnostic rate using combined molecular methods, (iii) investigate the presence of the difficult-to-culture zoonotic agents Coxiella burnetii, Chlamydia spp., and Leptospira spp. in bovine abortions in Denmark. Tissues from 162 aborted or stillborn bovine fetuses and placentas submitted for routine diagnostics were screened for pathogenic bacteria using 16S rDNA amplicon sequencing. Lesion association of fungal elements, as well as of selection of bacterial abortifacients, was assessed using specific FISH assays. The presence of Chlamydia spp. and chlamydia-like organisms was assessed using qPCR. The study focused on bacterial and fungal abortifacients, because Danish cattle is free from most viral abortifacients. The 16S rDNA amplicon sequencing–guided FISH approach was suitable for enhancing abortion diagnostics, i.e., the diagnostic rate for cases with tissue lesions (n = 115) was increased from 46 to 53% when compared to routine diagnostic methods. Identification of Bacillus licheniformis, Escherichia coli, and Trueperella pyogenes accounted for the majority of additional cases with an established etiology. No evidence for emerging or epizootic bacterial pathogens was found. The difficult-to-culture abortifacients were either not detected or not identified as abortifacients.

Prevalence of bovine abortion, calf mortality, and bovine viral diarrhea virus (BVDV) persistently infected calves among pastoral, peri-urban, and mixed-crop livestock farms in central and Northwest Ethiopia

BackgroundBovine Viral Diarrhea virus (BVDV) is one of important diseases of cattle worldwide causing economic losses to the cattle industry primarily due to increased premature culling and decreased reproductive performance. The virus can cross the placenta during early pregnancy and result in the birth of persistently infected (PI) calves that are efficient transmitters of BVDV and serving as the primary reservoirs for BVDV. Relatively few studies have focused on understanding BVDV seroprevalence, virus detection, genotyping and its distribution in Africa. Most BVDV research in Ethiopia has involved serologic surveys in adult cattle, rather than the identification of PI calves, despite their role in viral shedding and recurring infections. A cross-sectional study was undertaken in three different livestock production systems of Ethiopia with the objective to estimate the prevalence of bovine abortion, calf mortality, and BVDV persistently infected calves.ResultsEar notch samples (882) collected from calves in 349 households were tested for BVDV antigen using antigen capture enzyme-linked immunosorbent assay (ACE). All samples tested were negative for BVDV antigen. The overall animal level crude abortion and calf mortality prevalence were 4.0% (95% CI: 2.9–5.2) and 9.2% (95% CI: 7.7–11.0) respectively. The lower BVDV PI prevalence may be due to a lower effective contact rate between cattle reared in small-scale extensive production systems in Ethiopia.ConclusionsThis is the first report of BVDV Ag test in Ethiopia and no PI was detected in calves in the study areas. Since BVDV is a disease of great economic importance, this study finding must be interpreted with care since absence of evidence is not evidence of absence and even a single BVDV infected animal can serve as source of infection and contribute to the persistent spread of the virus. Greater attention needs to be given to screening for PI animals through testing large number of animals and culling positive animals. Hence, future research should focus on regions and production systems with high BVDV seroprevalence followed by antigen ELISA or BVDV real-time PCR to detect persistently infected and acutely viremic animals.

Spatial distance between sites of sampling associated with genetic variation among Neospora caninum in aborted bovine foetuses from northern Italy

BackgroundNeospora caninum, a coccidian protozoan, represents an important cause of bovine abortion. Available N. caninum strains show considerable variation in vitro and in vivo, including different virulence in cattle. To which extent sexual recombination, which is possible in the intestines of domestic dogs and closely related carnivores as definitive hosts, contributes to this variation is not clear yet.MethodsAborted bovine foetuses were collected between 2015 and early 2019 from Italian Holstein Friesian dairy herds suffering from reproductive problems. A total of 198 samples were collected from 165 intensive farms located in Lombardy, northern Italy. N. caninum samples were subjected to multilocus-microsatellite genotyping using ten previously established microsatellite markers. In addition to our own data, those from a recent study providing data on five markers from other northern Italian regions were included and analysed.ResultsOf the 55 samples finally subjected to genotyping, 35 were typed at all or 9 out of 10 loci and their individual multilocus-microsatellite genotype (MLMG) determined. Linear regression revealed a statistically significant association between the spatial distance of the sampling sites with the genetic distance of N. caninum MLMGs (P < 0.001). Including data from this and a previous North Italian study into eBURST analysis revealed that several of N. caninum MLMGs from northern Italy separate into four groups; most of the samples from Lombardy clustered in one of these groups. Principle component analysis revealed similar clusters and confirmed MLMG groups identified by eBURST. Variations observed between MLMGs were not equally distributed over all loci, but predominantly observed in MS7, MS6A, or MS10.ConclusionsOur findings confirm the concept of local N. caninum subpopulations. The geographic distance of sampling was associated with the genetic distance as determined by microsatellite typing. Results suggest that multi-parental recombination in N. caninum is a rare event, but does not exclude uniparental mating. More comprehensive studies on microsatellites in N. caninum and related species like Toxoplasma gondii should be undertaken, not only to improve genotyping capabilities, but also to understand possible functions of these regions in the genomes of these parasites.Graphical

Infectious bovine abortions: observations from an organized dairy herd.

Abortions in dairy animals can be caused by several infectious agents. Identification of the actual causal agent(s) is important for formulating suitable control strategies. A 3-year (2016-2018) longitudinal study was conducted in a dairy farm following an abortion storm in the mid- to late gestations. The investigation focused on the seven major infectious abortifacient in cattle, viz. bovine alphaherpesvirus-1 (BoHV-1), bovine viral diarrhoea virus (BVDV), Neospora caninum, Brucella abortus, Coxiella burnetii, Leptospira Hardjo, and Listeria monocytogenes. High seroprevalence was observed for BVDV (79.4%), Leptospira (70.5%), BoHV-1 (53.5%), and Brucella (45.0%) at the beginning of the investigation (August 2016). The incidence proportion increased for BVDV, Leptospira, and Brucella in the following years of the investigation. A strong association of Brucella seropositivity with history of abortion (OR = 3.27) was recorded. Incidence of BoHV-1 reduced during the period of study coincident with systematic IBR inactivated marker vaccination of the herd. Sixty-four abortion cases were investigated for the identification of causative agent(s) by microbial culture, serological (ELISA), and molecular detection (PCR/ real-time PCR). Antibodies to BVDV, Brucella, BoHV-1, Leptospira, Neospora, and Coxiella were detected in 63, 61, 56, 35, 5, and 6 aborting cattle, respectively. Real-time PCR/PCR of clinical specimens detected DNA of Brucella, BoHV-1, Coxiella, Leptospira, and Listeria in 34, 13, 12, 9, and 4 abortion cases, respectively. BVDV and Neospora were not detected in any specimen samples. Brucella abortus isolated from the farm was determined as ST1 by multi-locus sequence typing (MLST). DNA of multiple agents were detected in 21 of the 64 cases (43.75%). Overall, the data suggests, Brucella was the major causative agent, although multiple causative agents circulated in the farm.

Genetic Characterization of Mycoplasma. bovis, L. moncytogenes and Brucella species Recovered from Bovine Abortion

Genetic Characterization of Mycoplasma. bovis, L. moncytogenes and Brucella species Recovered from Bovine Abortion

Campylobacter fetus in Abomasal Fluid from Spontaneously Aborted Bovine and Ovine Fetuses

Background: Pregnancy losses are a major concern in livestock industry due to their economic impact on producers. Campylobacter fetus subspecies fetus (Cff) and C. fetus subspecies venerealis (Cfv) are directly related to reproductive failures in ruminants. Cff colonizes the gastrointestinal tract of a wide range of hosts leading to abortion, while Cfv is restricted to genital tract being generally associated to infertility in bovine. Considering the great economic losses related to campylobacteriosis in cattle and ovine herds, this study aims to investigate the occurrence of C. fetus, considering Cff and Cfv subspecies, in bovine and ovine spontaneously aborted fetuses in state of Rio Grande do Sul, Brazil.Materials, Methods &amp; Results: In this study, samples of abomasal fluid collected from 30 spontaneously aborted bovine (n = 18) and ovine (n = 12) fetuses were investigated for the detection of Campylobacter fetus throughout conventional PCR. Positive fetuses for C. fetus presence were further analyzed by molecular assays for Cff and Cfv detection, in order to determine subspecies identification. When available, samples of the main organs of the thoracic and abdominal cavities, as well as the brain, skeletal muscle, eyelid, skin, and placenta were collected for further histopathological analyses and bacterial culture, aiming to assess the presence of infection lesions and pathogens in those sites, respectively. Additionally, RT-qPCR assays were also performed for the detection of ruminant pestivirus, in order to detect bovine viral diarrhea cases. Throughout the present methodology, C. fetus was detected in the abomasal fluid samples of 2 bovine fetuses, being both identified as Cfv subspecies by PCR. Histopathological analyses demonstrated that macroscopic and microscopic changes found in the Cfv-positive animals were not either specific or directly related to Campylobacter infections. Moreover, no significant bacterial growth was observed in microbiological culture from the collected tissues, and both fetuses were negative for ruminant pestivirus. Differently, there was no detection of C. fetus in any of the analyzed ovine fetuses.Discussion: Considering that abortion diagnosis rates reported in cattle and sheep industry are highly variable among the published studies, and that abortion diagnoses are commonly inconclusive due to difficulties in sampling methodology and inadequate identification of the pathogen involved, it is important to investigate the etiological causes of abortion the herds for better understanding the causes of pregnancy issues and monitoring their occurrence. In addition, the absence of pathognomonic lesions in the tissues investigated in the histopathological analyses observed in this study strongly suggests that well-known etiological agents commonly associated to abortion, such as Leptospira spp., Toxoplasma spp., Chlamydia spp. and Neospora caninum, are unlikely to be the cause of infection of the analyzed fetuses. Taking this into account, the presence of C. fetus in the abomasal fluid samples from two bovine fetuses demonstrated in the present study suggests the possible association of Cfv not only with infertility, but also with cases of bovine abortion, highlighting the importance of investigating unusual causal agents of abortions in sheep and cattle. Overall, an adequate diagnosis is essential for establishing better prevention strategies to avoid the circulation of abortion-related infectious agents in the herds.Keywords: campylobacteriosis, molecular diagnosis, venereal disease, abortion, reproductive disease.

Variation in the Multivariate Relationships Between Serum Amyloid, Haptoglobin and Biochemical Profiles of Aborted and Non-Aborted Cows

Because the time in which gestational loss or bovine abortion occurs is associated with multifactorial causes, the objective of this study was to assess multivariate relationships between the biochemical profile and acute phase proteins in aborted cows. We performed monitoring and monthly blood sampling during gestation in 140 dairy cows. A total of 18 cows aborted, which were considered ‘cases’; retrospectively, we used samples from 29 ‘control’ cows, which were taken at the same time. Thus, we compared multivariate relationships between the two groups, to evaluate variables potentially associated with cow abortion. We classified abortion as either infectious (due to DVB, leptospirosis or neosporosis) or non-infectious (protein or energetic imbalance evinced in metabolic profile). Of the 18 cows that aborted, there were 6 with evidence of positivity or seroconversion to the analysed diseases, 13 with evidence of metabolic protein imbalances, 7 with evidence of energy imbalances and 10 of them with combined situations. Case and control cows have significant differences (p&lt;0.05) between their Principal Component Analysis (PCA), indicating that the two groups have different multivariate relationships. In aborted cows we identified a grouping of variables referring to protein metabolism asFibrinogen, Serum amyloid A, haptoglobin and titres for neosporosis and bovine viral diarrhoea virus; separated from others referring to energy and protein imbalance ashigh beta-hydroxy-butyrate, low cholesterol, low total protein. However, serum amyloid A and haptoglobin were not increased in aborted cows with clues of infectious diseases such as bovine viral diarrhoea virus, leptospirosis or neosporosis. In this way, it is evident that when the causality of abortions in a herd is studied, not only is it necessary to look for infectious diseases, but non-infectious causes can also be found occurring in parallel way. Serum amyloid A could be a candidate biomarker to differentiate a non-infectious abortion.

Investigation of Neospora caninum Seroprevalence and Association with Reproductive Problems in Cows in Burdur Province of Turkey.

Background:An apicomplexan protozoon Neospora caninum, causative agent of neosporosis, is recognized as one of the most common and important cause of sporadic and endemic bovine abortion and reduced reproductivity in dairy and beef cattle worldwide. The present study was conducted to investigate the relationship between N. caninum seroprevalence and infertility problems in 400 cows in Burdur, city, Turkey.Methods:Blood samples were collected from vena jugularis into sterile serum tubes from 49 aborted, 58 infertil, 48 pregnant and 245 healthy cows for the findings of reproductive anamnesis during a period of March 2010 to March 2011. Sera samples were analyzed by competitive ELISA kit.Results:The seroprevalences were 7.7%, 6.4% and 4.2% in 2–4, ≤2 and ≥4 age groups respectively and no statistically significance observed between age groups. Seropositivity rates were 5.7%, 5.1%, 4.5%, 3.6% in Holstein, Montofon, cross-breeds and Simental breeds respectively. Seroprevalence differences was not statistically significant among cattle breeds. Antibodies to N. caninum were found in rates of 16.3%, 6.9%, 6.3%, 2.4% in aborted, infertile, pregnant and healthy cows respectively and there was a significant difference (P<0.01) between aborted and healthy animals. Seroprevalences were Yeşilova 10%, Gölhisar and Ağlasun 8%, Bucak, Çavdır and Kemer 4%, Karamanlı and Burdur Centrum 2%, according to districts.Conclusion:The seroprevalence of N. caninum was revealed in Burdur region. It was emphasized that N. caninum infection should not be ignored in reproductive problems, especially in abortion cases.

Microsatellite genotyping reveals extensive genetic diversity in bovine Neospora caninum from the humid Pampa region in Argentina.

Neospora caninum is an apicomplexan protozoan and a major cause of abortion in cattle worldwide. In the Argentinian Humid Pampa, bovine neosporosis causes severe economic losses. Despite this, information on the genetic structure of N. caninum in this region is limited. Therefore, this study aimed to genetically characterize N. caninum isolates associated with bovine abortion in the Humid Pampa region. For this purpose, spontaneous bovine fetal tissues submitted for diagnosis to the Veterinary Diagnostic Service at INTA Balcarce during 2008-2019 were assessed by PCR, indirect fluorescent antibody test (IFAT), and histologic analysis. PCR-positive samples were tested by multilocus microsatellite genotyping (MLGs) using 9 microsatellite markers. Thirty-one different genotypes were identified from 32 samples with at least seven markers. Argentinian MLGs were grouped into two clonal clusters when analyzed using eBURST network and principal coordinate analysis. No segregation based on the year of collection, animal biotype, or geographic origin was observed. In addition, the presence of linkage disequilibrium supported the clonal propagation of Argentinian MLGs. One Argentinian subpopulation was associated with isolates from Spain, Uruguay, Brazil, and Mexico, and the other one was linked to isolates from Scotland, Spain, and Germany. These findings reveal the presence of two clonal subpopulations of N. caninum in the Humid Pampa.

Improvement of Leptospira spp. diagnosis in aborted bovine fetuses by qPCR

Leptospirosis is a disease with major economic impact on livestock industry. The objective of this work was to determine the presence of Leptospira spp. DNA by qPCR in bovine fetuses with presumptive diagnosis of leptospirosis as the cause of abortion. Leptospira spp. DNA was detected by qPCR in 11 out of 34 fetuses. These specimens (10/11) had histopathological findings in hepatic and/or renal tissues compatible with leptospirosis. qPCR detection rate (32.4 %) was higher compared with direct immuno-fluorescence antibody test (DFAT) (11.8 %). The concordance coefficient between both techniques was 0.44. qPCR is a rapid and sensitive technique for the diagnosis of leptospirosis and improved the detection rate in fetal tissues compared with DFAT. Implementation of molecular techniques may increase the accurate detection of leptospirosis as a cause of bovine abortion allowing the application of rapid therapeutic and prophylactics measures in order to reduce the impact of this zoonotic disease.

Sero-prevalence and Risk Factor Analysis of BoHV-1 in Bovines in Haryana State of India

Background: Bovine herpesvirus-1 (BoHV-1) is an important pathogen of cattle and buffaloes associated with various clinical conditions including infectious bovine rhinotracheitis (IBR) and abortion. To know the status of BoHV-1, a cross-sectional serological study was conducted with the objectives of estimating the apparent prevalence of BoHV-1 and potential risk factor among unorganized cattle and buffalo herds. Method: A total of 490 serum samples were collected from cattle and buffaloes from all twenty two (22) districts of Haryana from unorganised herd randomly and tested for antibodies against BoHV-1 using ELISA. Result: The overall percent sero-prevalence of BoHV-1 was observed as 48.78% however the species wise sero-prevalence was 37.77% in cattle and 62.27% in buffaloes. The overall sero-prevalence was significantly (p less than 0.05) associated with species, zone and age of animals. The likelihood of BoHV-1 was significantly higher (2.72 times) in buffaloes (Odds ratio (OR) =2.72, 95% Confidence Interval (CI):1.86; 3.98) than in cattle (OR=1). Eastern zone of the state showed higher (1.52 times, 95% CI: 1.03, 2.26) likelihood of BoHV-1 as compared to western zone (OR=1.00).The aged animals with age³6.5 years (2.96 times), followed by 2.5-4.5 years (2.44 times) and 4.5-6.5 years (1.68 times) showed higher likelihood than younger animals (Age less than 2.5 years). Further, it can be concluded that BoHV1 is circulating among livestock population in the state.

Seroprevalence of Neospora caninum infection in cattle from Pereira, Colombia ⋆.

There are over 25.6 million cattle heads in Colombia being the fourth-highest herd in Latin America. This study aimed to describe the seroprevalence of Neospora caninum in cattle from 25 rural farms at Pereira municipality, Risaralda Department, which has a total bovine population of 43,508 animals. A cross-sectional observational study was performed in beef and dairy herds during 2017-2018. A total of 325 animals in 25 herds, with 13 animals per herd, were sampled. A commercial competitive ELISA (cELISA) kit was used to detect N. caninum antibodies. Associated risk factors were analyzed in two different levels: individual animals and herds. For all the independent variables, chi-square (χ2) and Fisher tests were used to assess associations and significance. The overall estimated seropositivity was 20.6% (95%CI 16.2%-25.0%). The seroprevalence by herds was 92.0% ranging from 0.0% to 46.2%. Noteworthy, beef herds had significantly (p=0.0107) higher seropositivity (50% of them above 35% of seropositivity) compared with those for milk purposes (4.8%) (OR=20.0; 95%CI 1.2-331.0). Other risk factors were not significantly (p≥0.05) associated with N. caninum seropositivity. Bovine neosporosis may be associated with abortions in both beef and dairy Colombian farms. More studies about the epidemiology, associated factors and consequences, as well as on tools for better diagnosis of bovine abortion, including histopathology and other ancillary tests, should be performed.

Laboratory Diagnosis of Bovine Abortions Caused by Non-Maintenance Pathogenic Leptospira spp.: Necropsy, Serology and Molecular Study Out of a Belgian Experience.

Bovine leptospirosis is a bacterial zoonotic disease caused by pathogenic Leptospira spp. The pathology and epidemiology of this infection are influenced by the numerous existing serovars and their adaptation to specific hosts. Infections by host-maintained serovars such as Hardjo are well documented, unlike those from the incidental ones. In July 2014, an emerging phenomenon of an increased incidence of icteric abortions associated with leptospiral infection occurred in southern Belgium. First-line serological analyses targeting cattle-adapted serovars failed at initial diagnosis. This study provides a comprehensive description of laboratory findings—at the level of necropsy, serology and molecular diagnosis—regarding icteric and non-icteric abortions (n = 116) recorded during this time (years 2014–2015) and associated with incidental infection by serovars such as Grippotyphosa, Australis and Icterohaemorrhagiae. Based on these tests, a diagnostic pathway is proposed for these types of infection in cattle to establish an affordable but accurate diagnosis in the future. These investigations add insights into the understanding of the pathogenesis of bovine leptospirosis associated with serovars classically described as non-maintenance.

Causes of abortion in dairy cows in Uruguay

ABSTRACT: A case series study was conducted to determine the frequency of causes of abortion in dairy cattle in Uruguay. The sample size of 102 cases was composed of 53 fetuses, 35 fetuses with placentas, and 14 placentas without an associated fetus. All cases underwent gross and microscopic pathologic examinations as well as microbiological and serological testing. The etiology was determined in 54 (53%) of cases, 51 of which were caused by infectious agents. Within the observed 102 cases, 30 (29%) were caused by Neospora caninum, six (6%) by Coxiella burnetii and two (2%) by Campylobacter fetus subsp. venerealis. Bovine Parainfluenza-3 virus and Salmonella enterica serovar Newport caused one abortion each. Opportunistic bacteria (Escherichia coli, Streptococcus sp., Staphylococcus sp., Mannheimia sp., Trueperella pyogenes, and Providencia stuartii) were associated with 11 abortions. In two cases the fetal death was attributed to dystocia, and in one case the fetus had a congenital mesothelioma. Bovine viral diarrhea virus (BVDV) infection was identified in three fetuses; two of which were co-infected with and had typical lesions of N. caninum. No lesions were observed in the other fetus infected by BVDV. Leptospira interrogans was identified in one fetus without lesions. Despite the relatively low overall success rate in establishing an etiological diagnosis in cases of abortion in cattle, a systemic workup of bovine abortion is necessary to establish prevention and control strategies. This also facilitates monitoring and surveillance of reproductive diseases in dairy cattle, some of which represent a risk to public health.

Bovine abortion, stillbirth and neonatal death associated with Babesia bovis and Anaplasma sp. infections in southern Brazil

Anaplasmosis and babesiosis are tick-borne diseases widely disseminated in cattle herds in many parts of the world. These diseases represent important causes of death and economic losses in several countries, including Brazil, and are characterized by hemolytic disease and anemia. Animals of all ages may be affected. Although transplacental infections are known to occur, abortion, stillbirth and neonatal death directly associated with Anaplasma marginale and especially Babesia spp. infections have rarely been documented in cattle. The objective of the present study is to describe the pathological and molecular findings of two cases of bovine abortion, two cases of stillbirth and two cases of neonatal death associated with intrauterine anaplasmosis and/or babesiosis in southern Brazil. All cases occurred in beef farms in the state of Rio Grande do Sul, between 2017 and 2019. Angus and crossbred calves were affected. At the necropsy, the main gross lesions observed included different degrees of splenomegaly, enlarged and yellow liver, thick and grumous bile, pallor or jaundice of mucous membranes and carcass, and dark kidneys. Four calves also presented cherry-pink discoloration of the central nervous system. Cytological slides enabled the observation of intraerythrocytic organisms consistent with Babesia bovis (3/6) and A. marginale (2/6). Through PCR assays, it was possible to detect three cases of Babesia sp. infection alone, and one case of Anaplasma sp. infection alone. Co-infections with Anaplasma sp. and Babesia sp. were detected in two cases. These findings reaffirm that anaplasmosis and babesiosis should be considered as an important differential diagnosis of fetal loss, stillbirth and neonatal death in cattle in areas where these diseases occur.

Leptospira Interrogans Serogroup Sejroe Serovar Hardjo in Aborting Cows: Two Herd Cases in Sicily (Italy).

IntroductionThe aim of this study was to present two outbreaks of bovine abortion due to Leptospira infection in cattle herds located in the northern part of Sicily (Italy). The animals were positive for Leptospira interrogans serogroup Sejroe serovar Hardjo in a microscopic agglutination test (MAT).Material and MethodsA total of 23 Charolaise cows (farm A) and 75 Limousine bulls and Cinisara and Modicana cows (farm B) were enrolled in this study. The blood samples were collected from all subjects at the following time points: before a cycle of intramuscular treatment with oxytetracycline dihydrate (T0), after 5–6 weeks from the treatment (T1), and every 10 weeks until seronegativisation (T2 in Farm A and T3 in Farm B). A serological test (MAT) was used for the diagnosis of leptospirosis.ResultsTwo samples from farm A (2/23) and 29 samples from farm B (29/75) were positive to Leptospira interrogans, serogroup Sejroe, serovar Hardjo in the MAT. Leptospira spp. DNA was detected by real-time PCR in the urine sample of one positive cow on farm A, and in placenta and brain samples belonging to one aborted foetus on farm B.ConclusionIt is important to use serological and molecular diagnostic techniques complementarily to identify infected individuals.

Isolation and genetic characterization of Neospora caninum from naturally infected sheep

Neospora caninum is considered one of the main causes of abortion in cattle but can also cause abortion in sheep. There is limited knowledge of the N. caninum population infecting sheep, and only one N. caninum isolate from a pregnant sheep from Japan has been reported. This study describes the in vitro isolation and genetic characterization of two new sheep isolates of N. caninum implicated in ovine reproductive failure. We used IFN-γ-knockout mice inoculated with PCR-positive brain homogenates from two clinically healthy but congenitally infected lambs at 4.5 months of age for parasite isolation. The lambs were born to dams from a sheep farm that had experienced pregnancy failure caused by N. caninum in successive generations. Tachyzoites were microscopically visualized in peritoneal flushes from all inoculated mice and were also observed in MARC-145 cell cultures within one week after inoculation with peritoneal flushes. Two N. caninum isolates, Nc-Spain11 and Nc-Spain12, were obtained from each lamb. The genotyping of the Nc-Spain11 and Nc-Spain12 isolates based on 9 microsatellite markers showed identical multilocus genotype (MLG). Comparison between a previous N. caninum genotype dataset including 80 MLGs from Argentinean, Spanish, Mexican, German and Scottish bovine isolates and the Japanese sheep isolate showed that the Nc-Spain11 and Nc-Spain12 MLG was unique and differed from the other MLGs. eBURST analyses showed that the Nc-Spain11 and Nc-Spain12 MLG was genetically clustered with other bovine MLGs and one ovine MLG, and the nearest genetic relationship was with an MLG from a bovine abortion collected in the same geographical area of Galicia.