A G3P[13] porcine group A rotavirus emerging in China is a reassortant and a natural recombinant in the VP4 gene

Group A rotaviruses (RVA s) are a major cause of serious intestinal disease in piglets. In this study, a novel pig strain was identified in a stool sample from China. The strain was designated RVA /Pig/China/LNCY /2016/G3P[13] and had a G3‐P[13]‐I5‐R1‐C1‐M1‐A8‐N1‐T1‐E1‐H1 genome. The viral protein 7 (VP 7) and non‐structural protein 4 (NSP 4) genes of RVA /Pig/China/LNCY /2016/G3P[13] were closely related to cogent genes of human RVA s, suggesting that a reassortment between pig and human strains had occurred. Recombination analysis showed that RVA /Pig/China/LNCY /2016/G3P[13] is a natural recombinant strain between the P[23] and P[7] RVA strains, and crossover points for recombination were found at nucleotides (nt) 456 and 804 of the VP 4 gene. Elucidating the biological characteristics of porcine rotavirus (PoRV ) will be helpful for further analyses of the epidemic characteristics of this virus. The results of this study provide valuable information for RVA recombination and evolution and will facilitate future investigations into the molecular pathogenesis of RVA s.     

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.     

First experimental proof of Rotavirus A (RVA) genotype G18P[17] inducing the clinical presentation of ‘young pigeon disease syndrome’ (YPDS) in domestic pigeons (Columba livia)

Young pigeon disease syndrome (YPDS) is characterized as a seasonally occurring, acute and primarily enteric medical condition of mainly juvenile domestic pigeons (Columba livia) with highly variable mortality reaching more than 50%. Although the syndrome has been known in Europe for almost three decades, its aetiology remains largely obscure. Recently, a previously unknown pigeon‐associated clade of Rotavirus A (RVA) genotype G18P[17] was detected in Europe and Australia in association with fatal diseases resembling YPDS. Here we show for the first time, that peroral inoculation of healthy juvenile homing pigeons with two genetically different cell culture isolates of RVA G18P[17] (10^6.3 foci‐forming units per bird) induces an acute and self‐limiting YPDS‐like disease in all infected birds. Clinical signs included regurgitation, diarrhoea, congested crops, anorexia and weight loss, as described for naturally RVA‐infected pigeons. In agreement with the original outbreaks, RVA isolate DR‐7 induced more pronounced clinical signs as compared to isolate DR‐5, indicating strain‐dependent virulence factors to contribute to variable disease outcomes observed in the field. All inoculated birds developed rotavirus‐reactive antibodies starting at seven days after inoculation. High levels of viral RNA and infectious virus were detectable in cloacal swabs and faecal samples already three days after inoculation. While shedding of infectious virus subsided within few days, moderate viral RNA levels were still detectable in cloacal swabs, faeces, and tissue samples at the end of the experiment three weeks after inoculation. Histopathological analysis at this time point revealed inflammatory lesions in spleens and livers of pigeons from both infected groups. In summary, we fulfilled Henle‐Koch's postulates and confirmed RVA G18P[17] as a primary cause of YPDS‐like diseases in domestic pigeons. By establishing an infection model, we provide a crucial tool for future research, such as identification of transmission routes and establishing vaccination regimes.     

Emergence of novel lineage of foot‐and‐mouth disease virus serotype Asia1 BD‐18 (G‐IX) in Bangladesh

Foot‐and‐mouth disease virus (FMDV) is a highly evolutionary divergent pathogen causing great economic havoc in many countries. Among its seven existing serotypes, Asia1 is the least divergent with a single topotype both genetically and antigenically. It is reported sporadically in Indian subcontinent and was classified under lineage G‐VIII. In 2018, serotype Asia1 re‐emerged in Bangladesh after 2013, along with circulation of a novel serotype Asia1 BD‐18 (G‐IX) lineage. VP1 phylogeny and sequence variation clearly demonstrated the novel strains which was estimated to have at least >5% nucleotide divergence with distinct clade formation. Also, the Bayesian phylogeographic inferences traced back to the origin time of lineage G‐IX in early 2017 and a possible origin in Bangladesh. Mutational analysis considering established eight lineages revealed that the virus strains belonged to lineage G‐IX contained a unique mutation at 44 position in the B‐C loop region of VP1. Inappropriate vaccination and inefficient outbreak surveillance possibly contributed to the current episode of emergence. Therefore, active surveillance and continued vigilance are essential to assess and timely detect the occurrence, extent and distribution of this novel Asia1 strains in Bangladesh and the neighbouring countries.     

High syphilis seropositivity in European brown hares (Lepus europaeus), Lower Saxony,Germany

The lagomorph‐infecting Treponema paraluisleporidarum is a close relative of the human syphilis‐bacterium Treponema pallidum. There is a paucity of information on the epidemiology of hare syphilis and its relationship to the rabbit‐ and human‐infecting treponemes that cause syphilis. In our study, we tested 734 serum samples from European brown hares (Lepus europaeus) collected between 2007 and 2019 in the federal state of Lower Saxony, Germany, for the presence of antibodies against T. paraluisleporidarum. Since T. paraluisleporidarum cross‐reacts with T. pallidum antigen, we used a commercially available T. pallidum‐particle agglutination (TP‐PA) assay to test for the presence of antibodies. A high seropositivity (n = 405/734) was detected. An additional 233 serum samples were retested using a fluorescent treponemal antibody absorption test to confirm the results of the TP‐PA assay. Our results show that infection is widespread in Lower Saxony and suggest a horizontal (sexual) transmission mode since adult hares show significantly higher seropositivity than subadults (odds ratio: 0.03 [95% CI 0.02–0.05], p < .0001). No difference was detected based on gender (odds ratio: 0.79 [95% Cl 0.58–1.07], p = .1283). Further studies are warranted to genetically characterize the T. paraluisleporidarum strains that infect wild hares.     

Detection and phylogenetic analysis of porcine bocaviruses carried by murine rodents and house shrews in China

Bocaparvovirus infections of humans and both wild and domestic animals have been widely reported around the world. In this study, we detected and genetically characterized porcine bocavirus (PBoV) carried by murine rodents (Rattus norvegicus, Rattus tanezumi, and Rattus losea) and house shrews (Suncus murinus) in China. Between May 2015 and May 2017, 496 murine rodents and 23 house shrews were captured in four Chinese provinces. Nested polymerase chain reaction was used to investigate the prevalence of PBoV in throat swab, faecal and serum samples. A total of 7.5% (39/519) throat swab samples, 60.5% (309/511) faecal samples, and 22.9% (52/227) serum samples were PBoV‐positive. The prevalence among R. norvegicus and R. tanezumi was higher than that among R. losea and house shrews. PBoV‐positive samples were found in all four provinces. Phylogenetic analysis based on partial viral capsid protein 1/2 (VP1/VP2) showed that sequences obtained in this study formed a novel group (PBoV G4). In addition, five near full‐length PBoV genomes (4,715–4,798 nt) were acquired. These genomes encoded two non‐structural proteins, NS1 (1,908 nt in four genomes and 1,923 nt in the remaining genome) and NP1 (600 nt), and the structural proteins, VP1/VP2 (1,851 nt). Phylogenetic analysis showed that PBoV G4 is distinct from rodent, human, and other bocaviruses. In conclusion, PBoV G4 prevalence was high among two common murine rodents in China, and the pathogenecity of PBoV G4 need to be further clarified.     

Isolation and Potential for Transmission of Mycobacterium bovis at Human–livestock–wildlife Interface of the Serengeti Ecosystem,Northern Tanzania

Mycobacterium bovis, the causative agent of bovine tuberculosis (bTB), is a multihost pathogen of public health and veterinary importance. We characterized the M. bovis isolated at the human–livestock–wildlife interface of the Serengeti ecosystem to determine the epidemiology and risk of cross‐species transmission between interacting hosts species. DNA was extracted from mycobacterial cultures obtained from sputum samples of 472 tuberculosis (TB) suspected patients and tissue samples from 606 livestock and wild animal species. M. bovis isolates were characterized using spoligotyping and Mycobacterial Interspersed Repetitive Units‐Variable Tandem Repeats (MIRU‐VNTR) on 24 loci. Only 5 M. bovis were isolated from the cultured samples. Spoligotyping results revealed that three M. bovis isolates from two buffaloes (Syncerus caffer) and 1 African civet (Civettictis civetta) belonged to SB0133 spoligotype. The two novel strains (AR1 and AR2) assigned as spoligotype SB2290 and SB2289, respectively, were identified from indigenous cattle (Bos indicus). No M. bovis was detected from patients with clinical signs consistent with TB. Of the 606 animal tissue specimens and sputa of 472 TB‐suspected patients 43 (7.09%) and 12 (2.9%), respectively, yielded non‐tuberculous mycobacteria (NTM), of which 20 isolates were M. intracellulare. No M. avium was identified. M. bovis isolates from wildlife had 45.2% and 96.8% spoligotype pattern agreement with AR1 and AR2 strains, respectively. This finding indicates that bTB infections in wild animals and cattle were epidemiologically related. Of the 24 MIRU‐VNTR loci, QUB 11b showed the highest discrimination among the M. bovis strains. The novel strains obtained in this study have not been previously reported in the area, but no clear evidence for recent cross‐species transmission of M. bovis was found between human, livestock and wild animals.     

The first report of animal genotypes of Cryptosporidium parvum in immunosuppressed and immunocompetent humans in Slovakia

The aim of our study was to determine species and genotypes of Cryptosporidium in patients suffering from immunosuppressive illnesses, but also in immunocompetent patients suffering from diarrhoea. A total of 80 samples of faeces were collected from both immunosuppressed and immunocompetent patients. The immunosuppressed patients (65 samples) — 35 adult patients (group A) and 30 children (group B) were hospitalized at the Clinic of Oncohemathology. Samples from immunocompetent humans (15 samples, group C) were taken from patients with clinical signs of acute diarrhoea. With the use of molecular methods targeting the 60 kDa glycoprotein (GP60) gene region, we have identified multiple genotypes of Cryptosporidium. parvum and Cryptosporidium. hominis in immunocompromised, but also in immunocompetent individuals (C. hominis IbA10G2, IeA12G3T3; C. parvum IIaA10G1R1, IIaA11G2R1, IIaA12G2R1, IIaA13G1R1, IIaA14G1R1, IIaA14G2R1, IIaA17G1R1 and IIaA18G1R1). This is the first report of the occurrence of genotypes IIaA10G1R1, IIa12G2R1 and IIaA18G1R1 in human hosts.     

Production and Diagnostic Application of a Purified,E. coli‐Expressed,Serological‐Specific Chicken Anaemia Virus Antigen VP3

The aim of this study was to evaluate the production of chicken anaemia virus VP3 protein in different Escherichia coli strains and to address the diagnostic application of purified E. coli‐expressed VP3 protein for the detection of chicken anaemia virus (CAV) infection and the development of an ELISA kit. Three E. coli strains, BL21, BL21 codonplus RP and BL21 pLysS, each harbouring a VP3 protein expressing plasmid, were investigated after induction to produce recombinant VP3 protein. After isopropyl‐β‐d ‐thiogalactoside (IPTG) induction, VP3 protein was successfully expressed in all three E. coli strains. The BL21 pLysS strain gave the best performance in terms of protein productivity and growth profile. In addition, the optimal culture temperature and IPTG concentration were found to be 0.25 mm and 20°C, respectively. Using Ni‐NTA‐purified VP3 protein as an ELISA coating antigen, the purified VP3 was shown to be highly antigenic and able to discriminate sera from chickens infected with CAV from those that were uninfected during an evaluation of CAV infection serodiagnosis. A VP3‐based ELISA demonstrated 100% (6/6 × 100%) specificity and sensitivities of 91.3% (21/23 × 100%) and 82.6% (19/23 × 100%) using cut‐off values of the mean plus 2 SD and the mean plus 3 SD, respectively.     

Characterization of transboundary foot‐and‐mouth disease viruses in Nigeria and Cameroon during 2016

Continuous surveillance for foot‐and‐mouth disease (FMD) in endemic settings such as West Africa is imperative to support improved local and regional control plans, with the long‐term goal of regional eradication. This paper describes the genetic characterization of FMD viruses (FMDV) obtained from outbreaks in Nigeria (n = 45) and Cameroon (n = 15) during 2016 and from archival samples (n = 3) retrieved from a 2014 outbreak in Nigeria. These viruses were analysed in the context of previously published FMDV sequences from the region. Four FMDV serotypes: O, A, SAT1 and SAT2, were detected. Phylogenetic analyses of the VP1 coding sequences indicate the continuity of FMDV serotype O East Africa‐3 (O/EA‐3), serotype A AFRICA genotype G‐IV (A/AFRICA/G‐IV) and serotype South African Territories (SAT) 2 lineage VII (SAT2/VII). The FMDV SAT1 topotype X (SAT1/X), which emerged in Nigeria in 2015, continued to be associated with outbreaks in the region during 2016, and SAT1 is reported for the first time from Cameroon. Additionally, a re‐emergence or re‐introduction of the serotype O West Africa (O/WA) topotype in Nigeria is described herein. Our findings indicate a consistent, pan‐serotypic relationship between FMDV strains detected in Cameroon and Nigeria. Additionally, FMDV strains from West Africa obtained in this study were genetically related to those occurring in East and North Africa. These phylogenetic relationships suggest that animal movements (pastoralism and/or trade) are important factors for virus spread across the African continent. These data provide critical baselines which are a necessary component of Stages 0 and 1 of the Progressive Control Pathway of FMD (PCP‐FMD). Specifically, characterizing the existing virus strains (risk) provides the basis for the comprehensive risk‐based control plan which is the requisite criteria for Nigeria's transition to Stage 2 of PCP‐FMD, and for coordinated regional control of FMD.     

Senecavirus A: An Emerging Vesicular Infection in Brazilian Pig Herds

Vesicular diseases are clinically and economically important infections that affect farm animals. North American studies have suggested that Senecavirus A infection might be associated with a vesicular disease in pigs known as porcine idiopathic vesicular disease (PIVD). In the beginning of 2015, outbreaks of porcine vesicular disease have occurred in six Brazilian states from three geographical regions. Official diagnostic tests were performed with negative results for classical vesicular diseases of compulsory reporting. This study investigated Senecavirus A infection in PIVD outbreaks in which other aetiological agents were ruled out. A primer set was designed to amplify a 542‐bp product size of VP3/VP1 region of Senecavirus A genome in RT‐PCR assay. Primer specificity was analysed in silico and in porcine biological specimens. For this, clinical specimens were collected from eight pig herds affected with PIVD, including vesicular fluid (n = 4) and swabs (n = 7) and scrapings of ruptured vesicles and ulcerative lesions (n = 5) from weaned and adult pigs. Clinically healthy animals (n = 52) of PIVD‐affected and non‐affected pig herds also were evaluated for Senecavirus A infection. The 16 samples from PIVD‐affected animals were positive for Senecavirus A in the RT‐PCR assay, while none of the clinically healthy pigs were detected with the virus. Sequencing analysis revealed high nucleotide (87.6–98.5%) and amino acid (95–99.4%) similarities to SVV‐01 prototype and other Senecavirus A strains from North American pigs. Primer set presented herein was suitable for molecular characterization of Senecavirus A. The results suggest that Senecavirus A was the aetiological agent of the vesicular disease outbreaks in the evaluated pig herds. This is the first study to report the Senecavirus A infection in clinically affected pigs outside of North America. Senecavirus A was considered a novel emerging pathogen associated with an important vesicular disease in Brazil.     

Identification and molecular characterization of Staphylococcus aureus and multi‐drug resistant MRSA from monkey faeces in China

Staphylococcus aureus is a commensal bacterium and an important opportunistic pathogen in humans and animals. The increase in multi‐drug resistant (MDR) strains of S. aureus is a growing concern due to their impact on animal health and potential for zoonotic transmission. Increasing evidence has shown that MRSA could be transmitted by faeces. The present study determined the prevalence, antibiotic resistance profile and genotypic characteristics of S. aureus isolated from monkey faecal samples in China. Thirty‐eight out of 145 (26.21%) macaque faecal samples were S. aureus positive, which eight (5.5%) isolates were identified as MRSA. Antimicrobial susceptibility tests showed that most of the S. aureus isolates were resistant to tetracycline (TE, 44.74%), followed by penicillin (P, 21.05%), cefoxitin (FOX, 21.05%) and ciprofloxacin (CIP, 18.42%). The predominant spa types were t13638 (44.74%) and t189 (13.16%), which are reported to be closely associated with human infections in China. All MRSA isolates belonged to the SCCmecV type, which six of MRSA isolates were ST3268, while the other two isolates belonged to ST4981. This study for the first time describes the prevalence of S. aureus and MRSA in monkey faeces in China, indicating that faeces could be a potential factor of transmitting S. aureus between humans and monkeys.     

Cryoprotectant effect of trehalose in extender on post‐thaw quality and in vivo fertility of water buffalo (Bubalus bubalis) bull spermatozoa

This study was designed to ascertain the cryoprotectant effects of different concentrations of trehalose [0 (T0), 25 (T25), 35 (T35), 45 (T45) mm ], egg yolk [20% (E20), 15% (E15) v/v] and glycerol [7% (G7), 5% (G5) v/v] in Tris‐citric acid‐based extender on post‐thaw quality and in vivo fertility of buffalo bull spermatozoa. Twenty‐five ejaculates were collected from five bulls and split into four parts. After that, the split ejaculates from each of the bull were diluted either in T0E20G7 (control) or T25E20G5 or T35E15G5 or T45E15G5 extender. Finally, the sperm suspension was frozen in 0.54‐ml French straws. Post‐thaw sperm total motility (%), progressive motility (%), rapid velocity (%), average path velocity (μm/s), straightline velocity (μm/s), curvilinear velocity (μm/s), linearity (%), plasma membrane and acrosome integrities (%) were higher (p < .05) in T45E15G5 extender as compared to other treatment groups and control. The fertility rate (56.8% versus 41.3%) was higher (p < .05) in buffaloes inseminated with semen doses cryopreserved in extender containing T45E15G5 combination of cryoprotectants than the control. In conclusion, addition of 45 mm trehalose along with 15% egg yolk and 5% glycerol in extender improves the post‐thaw quality and in vivo fertility of buffalo bull spermatozoa.     

Characterization of porcine sapelovirus isolated from Japanese swine with PLC/PRF/5 cells

Porcine sapelovirus (PSV ) is a causative agent of neurological disorders, fertility disorders and dermal lesions of swine. In this study, we isolated two PSV strains, Jpsv477 and Jpsv1315, from swine faecal specimens using a PLC /PRF /5 cell culture system. The PSV infection of PLC /PRF /5 cells induced a cytopathic effect (CPE ). Two types of virus particles with identical diameter (~35 nm) but different densities (1.300 and 1.285 g/cm³) were observed in the cell culture supernatants. Analysis of the entire genome sequence of Jpsv477 and Jpsv1315 revealed that both strains possess 7,558 nucleotides and the poly (A) tail and have a typical PSV genome organization consisting of a 5′ terminal untranslated region (5′UTR ), a large open reading frame (ORF ), and a 3′ terminal untranslated region (3′UTR ). The ORF encodes a single polyprotein that is subsequently processed into a leader protein (L), four structural proteins (VP 1, VP 2, VP 3 and VP 4) and seven functional proteins (2A, 2B, 2C, 3A, 3B, 3C and 3D). The structural proteins VP 1, VP 2, VP 3 and VP 4 have molecular masses of ~35, ~26, ~25 and ~6 kDa. The N‐terminal amino acid sequence analysis of VP 1, VP 2, VP 3 and VP 4 confirmed that the cleavage sites between VP 4 and VP 2, VP 2 and VP 3, and VP 3 and VP 1 are K/A, Q/G and Q/G, respectively. We further confirmed that HepG2/C3A, Vero E6 and primary green monkey kidney cells (PGMKC ) were also susceptible to PSV infection. The stability assay demonstrated that PSV was inactivated by heating at 60°C for 10 min or 65°C for 5 min. The virus also lost infectivity by incubation with 62.5 ppm of NaClO for 30 min.     

Different surgical techniques and L‐carnitine supplementation in an experimental varicocele model

We aimed to investigate the impact of various varicocelectomy techniques and/or L‐carnitine as an adjunct treatment, following the emergence of oxidative stress, on the expression levels of SCF/c‐kit signalling pathways in spermatogenesis. Forty‐two rats were divided into seven groups: group 1 (G1) control; group 2 (G2) sham; group 3 (G3) varicocele; group 4 (G4) varicocele + varicocelectomy with testicular nonartery sparing; group 5 (G5) same as G4 but with artery sparing; group 6 (G6) same as G4 but with L‐carnitine and group 7 (G7) same as G5 with L‐carnitine. mRNA expression levels of SCF and c‐kit were measured quantitatively using real‐time polymerase chain reaction. CASP‐3 activity at protein level was determined, and histological evaluation was performed. mRNA expression level of SCF increased in G6 as compared to control group (3.52‐folds change; P = 0.035), whereas mRNA expression level of c‐kit gene remained the same. We found that in the left testis of G6 group, mRNA expression level of SCF increased 2.2‐folds in comparison with the right testis (P < 0.05). There were no statistically significant differences in the CASP‐3 protein expression levels between the control and other groups. When Cosentino Score analyses of immunostaining were conducted, we observed no significant differences among groups. Spermatogenic failure could be primarily due to a sertoli cell dysfunction. Although surgical treatment has been the best option for management of varicocele, auxiliary agents like L‐carnitine may be considered as supportive treatment regimes in addition to conventional surgical treatments.     

Tuberculosis Caused by Mycobacterium orygis in Dairy Cattle and Captured Monkeys in Bangladesh: a New Scenario of Tuberculosis in South Asia

Mycobacterium orygis, commonly known as the oryx bacillus and a newly proposed Mycobacterium tuberculosis complex subspecies, was isolated from 18 cattle in a dairy farm and two captured rhesus monkeys in a zoo in Bangladesh. All the infected animals had tuberculosis lesions in their lungs, suggesting transmission and infection with M. orygis by an airborne route. The 20 isolates were analysed using a range of conventional and molecular typing methods, and RD‐deletion typing and sequencing of selected genes confirmed the isolates as M. orygis. Multiple‐locus variable‐number tandem repeat analysis (MLVA) allowed the isolates to be divided into three clusters based on the relatedness of their MLVA profiles. The two monkey isolates shared the same MLVA pattern with 15 of the cattle isolates, whereas the remaining three cattle isolates had different patterns, even though the latter animals had been kept in the same dairy farm. The diversity observed among isolates may suggest the bacteria have been established in this area for a long period. This study along with other recent findings that report the detection of M. orygis from animals as well as humans originating from South Asia potentially indicate endemic distribution of M. orygis in South Asia.     

Short Report: Identification of a Potential Marker of Anaplasma Phagocytophilum Associated with Cattle Abortion

Anaplasma phagocytophilum is a tick‐borne pathogen that causes tick‐borne fever in domestic ruminants. Tick‐borne fever is characterized by diverse symptoms and occasionally causes abortions in domestic ruminants, resulting in significant economic impact. However, in France, the potential frequency of A. phagocytophilum‐related abortions is unknown, and thus, it remains difficult to estimate the full extent of its economic impact. This gap in our knowledge is likely explained, at least in part, by the absence of suitable and specific tools capable of detecting A. phagocytophilum associated with abortion. Our objective was to identify a genetic marker able to differentiate A. phagocytophilum strains isolated from domestic ruminants that had aborted compared to those that had not. Thus, we typed a total of 123 A. phagocytophilum obtained from cattle, of which 25 originated from cows that had aborted, via multiple‐locus variable‐number tandem repeat (VNTR) analysis. These included 56 new A. phagocytophilum samples and 67 previously published A. phagocytophilum samples. A multivariate logistic model demonstrated that the triple‐repeat allele of the APV‐A VNTR was statistically more frequent in A. phagocytophilum from cattle that had aborted, compared to A. phagocytophilum from cattle that had not aborted (P = 0.03), while controlling for any regional effects (P < 0.0001). For four other VNTR, there were no statistical associations between specific alleles and abortion. The APV‐A triple‐repeat VNTR allele could thus act as a marker of A. phagocytophilum involved in abortions. If this hypothesis is confirmed in additional samples from other regions, this marker could then be utilized in the development of A. phagocytophilum abortive strain surveillance measures.     

Molecular epidemiology of Leptospira noguchii reveals important insights into a One Health context

Leptospirosis presents a complex and dynamic epidemiology. Bovine leptospirosis has been described as a major infectious disease impairing reproductive efficiency. Although infections by Leptospira interrogans, L. santarosai and L. borgpetersenii are frequently reported in cattle, the presence of L. noguchii in these animals should not be neglected. In this study, we describe serological (MAT) and molecular characterization (rrs and secY gene sequencing, multilocus sequence typing [MLST] and pulsed‐field gel electrophoresis [PFGE]) of eight L. noguchii strains obtained from slaughtered cows. Intraspecific genetic diversity was evaluated, and haplotype networks were constructed based on hosts and geographical localizations. Strains were characterized as belonging to serogroups Australis, Autumnalis and Panama, and molecular characterization showed a high heterogeneity of these strains. Ten different STs were found (including nine new STs and 39 novel alleles) as well as nine different pulsotypes. Two clonal complexes were found. Phylogenetic trees based on secY locus and concatenated MLST loci showed two main clusters, with sequences from the present study included in the first. In general, there was no relationship between the geographical origin and the secY phylogenetic clusters, as well as between secY phylogenetic clusters and serogroups. Molecular diversity indexes confirmed a high variability (H > 0.8). This high intraspecific variation observed may be related to differences in virulence, pathogenicity and antigenicity or even adaptability of the strains. In addition, haplotype networks clearly demonstrated the circulation of genotypes between humans and animals, confirming the zoonotic potential. The present study provides relevant data for the study of leptospirosis in the One Health context, where human, animal and environmental health is closely connected.