Intensively Kept Pigs Pre-disposed to Chlamydial Associated Conjunctivitis

In the present study, ocular chlamydial infections in pigs that originate from two different farming systems were investigated. In particular, the aim was to test pigs with and without clinical ocular symptoms for the presence of Chlamydiaceae and for linked infections with Acanthamoebae spp. possibly acting as vectors for Chlamydia or Chlamydia-like organisms. In a total of 181 pigs, 102 from Germany (GER), representing the intensively kept animals and 79 from Switzerland (CH), which were kept extensively, were screened for the presence of different pathogens by PCR, including a new Chlamydiaceae-specific intergenic spacer rRNA gene PCR. Additionally, results of clinical examination and cytology were compared between the symptomatic and asymptomatic pigs of the two groups. Ocular symptomatic pigs showed a high prevalence of Chlamydia suis in both groups: CH 79%, GER 90%. Only 23% asymptomatic pigs from CH, but 88% asymptomatic pigs from GER were positive for C. suis by PCR. DNA of Chlamydia-like organisms were detected in 19% CH, but only in 2% GER pigs, whereas only 4% CH and 1% GER pigs were also positive for Acanthamoebae spp. A co-infection of Acanthamoebae spp. and C. suis was present in only 3% of the CH but 28% of the GER pigs. In general, the intensively kept pigs in our study seemed to be pre-disposed to ocular chlamydial infection and associated conjunctivitis. Infections with Chlamydia-like organisms alone and in combination with Acanthamoebae played no role for clinical findings within the tested pig groups, whereas a co-infection of Acanthamoebae and C. suis was able to cause serious ocular manifestations in half of the cases of intensively kept pigs being positive for these microorganisms.     

Assessment of Chlamydia suis Infection in Pig Farmers

Chlamydia suis infections are endemic in domestic pigs in Europe and can lead to conjunctivitis, pneumonia, enteritis and reproductive failure. Currently, the knowledge on the zoonotic potential of C. suis is limited. Moreover, the last decades, porcine tetracycline resistant C. suis strains have been isolated, which interfere with treatment of chlamydial infections. In this study, the presence of C. suis was examined on nine Belgian pig farms, using Chlamydia culture and a C. suis specific real‐time PCR in both pigs and farmers. In addition to diagnosis for C. suis, the farmers' samples were examined using a Chlamydia trachomatis PCR. Additionally, the Chlamydia isolates were tested for the presence of the tet(C) resistance gene. C. DNA was demonstrated in pigs on all farms, and eight of nine farmers were positive in at least one anatomical site. None of the farmers tested positive for C. trachomatis. Chlamydia suis isolates were obtained from pigs of eight farms. Nine porcine C. suis isolates possessing a tet(C) gene were retrieved, originating from three farms. Moreover, C. suis isolates were identified in three human samples, including one pharyngeal and two rectal samples. These findings suggest further research on the zoonotic transfer of C. suis from pigs to humans is needed.     

Endemic and Emerging Chlamydial Infections of Animals and Their Zoonotic Implications

The Chlamydiae are a diverse group of obligate intracellular Gram‐negative bacteria that are known to infect a wide variety of host species and are responsible for a wide range of diseases in animals and man. Many of these organisms have been extensively characterized and their zoonotic implications recognized. Studies of human disease first provided evidence for the disease‐causing potential of Chlamydia‐related bacteria; however, there is now increasing evidence that a number of these organisms may also be the causative agents for a number of pathogenic conditions of livestock that had previously remained undiagnosed. The aim of this review is to draw together the evidence for the role of the newly emerging chlamydial infections in livestock disease, the current understanding of their roles in human disease and highlight the potential for zoonotic transmission.     

Association d’une pneumopathie à Chlamydia psittaci et d’une maladie de HortonA possible assocation between Chlamydiae psittaci infection and temporal arteritis.

Some arguments are in favor of the role of Chlamydia in the pathogenesis of atherosclerosis and some vasculitis. Illustrating this possible relation, we report the case of a patient developing consecutively a Chlamydia psittacci infection and a temporal arteritis. A 73-years-old woman, with no significant medical history, was hospitalized for constitutionnal symptoms. Three weeks before, she described fever and sore throat during 2 days. Since that, she remained exhausted and developped a mild intermittent limp of the jaws. Clinical examination was poor. A biological inflammatory syndrome was noticed. X-ray chest revealed bilateral interstitial opacities. The titer of anti-C. psittaci antibodies was significant (positive IgG at 1/ 2048). Soon after initiation of doxycyclin, a temporal arteritis biopsy performed, due to the persistance of clinical symptoms and high inflammatory syndrom, concluded to the diagnosis of temporal arteritis. Corticotherapy was added to antibiotherapy resulting in decreasing of inflammatory syndrom and improvement of general status of the patient. X-ray opacities decreased in 3 weeks. Serological control after 3 months showed a decrease of the titer of anti-C.psittacci antibodies to 1/256, confirming the initial diagnosis of Chlamydia pneumopathy. Our observation could constitute one more argument for the role of bacteria like Chlamydia in the pathogenesis of vascular diseases. Prospective seroepidemiologic and molecular biology studies could allow to clarify the association between Chlamydia infections and inflammatory vasculitis like temporal arteritis.     

Brucella microti‐like prevalence in French farms producing frogs

In the last 10 years, many atypical novel members of Brucella species have been reported, including several Brucella inopinata‐like strains in wild‐caught and “exotic” amphibians from various continents. In 2017, a strain of Brucella was isolated for the first time in animals from a French farm producing frogs—Pelophylax ridibundus—for human consumption and identified as B. microti‐like. Following this first isolation, investigations were performed in this farm as well as in the farm of the research unit that provided the domestic frog strain to estimate the prevalence of B. microti‐like infection and its presence in the surrounding environment. Farming practices were investigated and samples including frogs at different development stages, surface tank swabs, water, feed and soil were analysed by real‐time PCR and bacteriological methods. High B. microti‐like prevalence values (higher than 90%) were obtained in frog samples in the commercial farm, and its presence was highlighted in the environmental samples except feed. In the research unit farm, B. microti‐like species was also isolated and detected in frog and environmental samples. These results show that B. microti‐like organisms are able to colonize amphibians and persist in their environment. Its presence could constitute a possible risk for consumers and workers proving the importance of assessing the zoonotic and pathogenic potentials of these new and atypical Brucella species.     

Myxoma virus jumps species to the Iberian hare

The study of myxoma virus (MYXV) infections in the European rabbit (Oryctolagus cuniculus) has produced one of the most accepted host–pathogen evolutionary models. To date, myxomatosis has been limited to the European rabbit with sporadic reports in hares. However, reports of widespread mortalities in the Iberian hare (Lepus granatensis) with myxomatosis‐like clinical signs indicate a potential species jump has occurred. The presence of MYXV DNA was confirmed by PCR in 244 samples received from regional veterinary services, animal health laboratories, hunters or rangers over a 5‐month period. PCR analysis of 4 MYXV positive hare samples revealed a 2.8 kb insertion located within the M009 gene with respect to MYXV. The presence of this insertion was subsequently confirmed in 20 samples from 18 Spanish provinces. Sanger sequencing and subsequent analysis show that the insert contained 4 ORFs which are phylogenetically related to MYXV genes M060, M061, M064 and M065. The complete MYXV genome from hare tissue was sequenced using Ion torrent next‐generation technology and a summary of the data presented here. With the exception of the inserted region, the virus genome had no large scale modifications and 110 mutations with respect to the MYXV reference strain Lausanne were observed. The next phase in the evolution of MYXV has taken place as a host species jump from the European rabbit to the Iberian hare an occurrence which could have important effects on this naïve population.     

Detection of a range of genetically diverse chlamydiae in Australian domesticated and wild ungulates

Chlamydiae are globally widespread obligate intracellular bacteria, which several species are a well‐recognized threat to human and animal health. In Australia, the most successful chlamydial species are the infamous koala pathogen C. pecorum, and C. psittaci, an emerging pathogen associated with zoonotic events. Little is known about infections caused by other chlamydial organisms in Australian livestock or wildlife. Considering that these hosts can be encountered by humans at the animal/human interface, in this study, we investigated genetic diversity of chlamydial organisms infecting Australian domesticated and wild ungulates. A total of 185 samples from 129 domesticated (cattle, horses, sheep, and pigs) and 29 wild (deer) ungulate hosts were screened with C. pecorum and C. psittaci species‐specific assays, followed by a screen with pan‐Chlamydiales assay. Overall, chlamydial DNA was detected in 120/185 (65%) samples, including all ungulate hosts. Species‐specific assays further revealed that C. pecorum and C. psittaci DNA were detected in 27% (50/185) and 6% (11/185) of the samples, respectively, however from domesticated hosts only. A total of 46 “signature” 16S rRNA sequences were successfully resolved by sequencing and were used for phylogenetic analyses. Sequence analyses revealed that genetically diverse novel as well as traditional chlamydial organisms infect an expanded range of ungulate hosts in Australia. Detection of the C. psittaci and C. pecorum in livestock, and novel taxa infecting horses and deer raises questions about the genetic make‐up and pathogenic potential of these organisms, but also concerns about risks of spill‐over between livestock, humans, and native wildlife.     

Molecular prevalence of Chlamydia and Chlamydia‐like bacteria in Tunisian domestic ruminant farms and their influencing risk factors

Chlamydia and Chlamydia ‐like bacteria are well known to infect several organisms and may cause a wide range of diseases, particularly in ruminants. To gain insight into the prevalence and diversity of these intracellular bacteria, we applied a pan‐Chlamydiales real‐time PCR to 1,134 veterinary samples taken from 130 Tunisian ruminant herds. The true adjusted animal population‐level prevalence was 12.9% in cattle, against 8.7% in sheep. In addition, the true adjusted herd‐level prevalence of Chlamydiae was 80% in cattle and 25.5% in sheep. Chlamydiales from three family‐level lineages were detected indicating a high biodiversity of Chlamydiales in ruminant herds. Our results showed that Parachlamydia acanthamoebae could be responsible for bovine and ovine chlamydiosis in central‐eastern Tunisia. Multivariable logistic regression analysis at the animal population level indicated that strata and digestive disorders variables were the important risk factors of bovine and ovine chlamydiosis. However, origin and age variables were found to be associated with bovine and ovine chlamydiosis, respectively. At the herd level, risk factors for Chlamydia positivity were as follows: abortion and herd size for cattle against breeding system, cleaning frequency, quarantine, use of disinfectant and floor type for sheep. Paying attention to these risk factors will help improvement of control programs against this harmful zoonotic disease.     

Chlamydial prostatitis in dogs: An experimental study

Summary Ten adult male mongrel dogs were inoculated with Chlamydia trachomatis by injections of 10⁶ or 10⁷ organisms directly into the prostate. We were able to recover Chlamydia 3 to 7 days later in three of four dogs receiving injections of 10⁷ organisms. Eight of ten dogs developed detectable serum antibody to Chlamydia 14 to 68 days following inoculation. All dogs receiving 10⁷ Chlamydia and two of six dogs receiving 10⁶ organisms developed histological signs of prostatic hyperplasia. Thus, the dog may be a model for establishment of chlamydial prostatitis and may be used for further investigations of this disease.This project was suppored in part by the Veterans Administration     

Bovine anaplasmosis and tick‐borne pathogens in cattle of the Galapagos Islands

A cross‐sectional study was conducted to determine the species of Anaplasma spp. and estimate its prevalence in cattle of the three main cattle‐producing Galapagos Islands (Santa Cruz, San Cristóbal and Isabela) using indirect PCR assays, genetic sequencing and ELISA . Ticks were also collected from cattle and scanned for 47 tick‐borne pathogens in a 48 × 48 real‐time PCR chip. A mixed effects logistic regression was performed to identify potential risk factors explaining Anaplasma infection in cattle. A. phagocytophilum was not detected in any of the tested animals. Genetic sequencing allowed detection of A. platys ‐like strains in 11 (36.7%) of the 30 Anaplasma spp.‐positive samples analysed. A. marginale was widespread in the three islands with a global between‐herd prevalence of 100% [89; 100]_95%CI and a median within‐herd prevalence of 93%. A significant association was found between A. marginale infection and age with higher odds of being positive for adults (OR = 3.3 [1.2; 9.9]_{95% Bootstrap CI} ). All collected ticks were identified as Rhipicephalus microplus. A. marginale , Babesia bigemina , Borrelia theileri and Francisella ‐like endosymbiont were detected in tick pools. These results show that the Galapagos Islands are endemic for A. marginale .     

Leishmaniasis in cat shelters: A serological,molecular and entomological study

An epidemiological Leishmania spp. and entomological Phlebotomine sandflies survey was performed in cat shelters at leishmaniasis endemic area of Brazil. Blood and conjunctival swab (CS) samples were collected from 94 cats in two animal protection shelters. These samples were subjected to serological tests using the indirect immunofluorescence antibody test (IFAT) and indirect enzyme‐linked immunosorbent assay (ELISA) and to molecular test by polymerase chain reaction (PCR). In addition, a Phlebotomine sandflies survey was performed in the same shelters. The analyses revealed a positivity of 31.91% (30/94) through ELISA and 29.79% (28/94) through IFAT. The two serological tests showed a positive association with perfect agreement (k = 0.925). None of the cats were positive by Leishmania spp. DNA. One Lutzomyia (Lutzomyia) longipalpis male was found in one of the cat shelters. The results and the implications of our findings are discussed below.     

Rapid Spread of Schmallenberg Virus‐infected Biting Midges (Culicoides spp.) across Denmark in 2012

Detection of Schmallenberg virus RNA, using real‐time RT‐PCR, in biting midges (Culicoides spp.) caught at 48 locations in 2011 and four well‐separated farms during 2012 in Denmark, revealed a remarkably rapid spread of virus‐infected midges across the country. During 2012, some 213 pools of obsoletus group midges (10 specimens per pool) were examined, and of these, 35 of the 174 parous pools were Schmallenberg virus RNA positive and 11 of them were positive in the heads. Culicoides species‐specific PCRs identified both C. obsoletus and C. dewulfi as vectors of Schmallenberg virus.     

Presence of aerobic micro‐organisms and their influence on basic semen parameters in infertile men

Urogenital tract infections in males are one of the significant etiological factors in infertility. In this prospective study, 72 patients with abnormal semen parameters or any other symptoms of urogenital tract infection were examined. Semen analysis according to the WHO 2010 manual was performed together with microbial assessment: aerobic bacteria culture, Chlamydia antigen test, Candida culture, Ureaplasma and Mycoplasma‐specific culture. In total, 69.4% of semen samples were positive for at least one micro‐organism. Ureaplasma sp. was the most common micro‐organism found in 33% of semen samples of infertile patients with suspected male genital tract infection. The 2nd most common micro‐organisms were Enterococcus faecalis (12.5%) and Escherichia coli (12.5%), followed by Staphylococcus aureus (7%), Chlamydia trachomatis (7%) and Candida sp. (5.6%). Generally, bacteria were sensitive to at least one of the antibiotics tested. No statistically significant relationship was observed between the presence of aerobic micro‐organisms in semen and basic semen parameters: volume, pH, concentration, total count, motility, vitality and morphology.     

Evidence of Presence of Mycobacterium tuberculosis in Bovine Tissue Samples by Multiplex PCR: Possible Relevance to Reverse Zoonosis

Bovine tuberculosis, caused by Mycobacterium bovis, remains one of the most important zoonotic health concerns worldwide. The transmission of Mycobacterium tuberculosis from humans to animals also occurs especially in countries where there is close interaction of humans with the animals. In the present study, thirty bovine lung tissue autopsy samples from an organized dairy farm located in North India were screened for the presence of Mycobacterium tuberculosis complex by smear microscopy, histopathological findings and PCR. Differential diagnosis of M. tuberculosis and M. bovis was made based on the deletion of mce‐3 operon in M. bovis. The present study found eight of these samples positive for M. tuberculosis by multiplex PCR. Sequencing was performed on two PCR‐positive representative samples and on annotation, and BLAST analysis confirmed the presence of gene fragment specific to Mycobacterium tuberculosis. The presence of M. tuberculosis in all the positive samples raises the possibility of human‐to‐cattle transmission and possible adaptation of this organism in bovine tissues. This study accentuates the importance of screening and differential diagnosis of Mycobacterium tuberculosis complex in humans and livestock for adopting effective TB control and eradication programmes.     

Carrier Status of Leptospirosis Among Cattle in Sri Lanka: A Zoonotic Threat to Public Health

Leptospirosis is a zoonotic disease of global importance and one of the notifiable diseases in Sri Lanka. Recent studies on human leptospirosis have suggested that the cattle could be one of the important reservoirs for human infection in the country. However, there is a dearth of local information on bovine leptospirosis, including its implications for human transmission. Thus, this study attempted to determine the carrier status of pathogenic Leptospira spp in cattle in Sri Lanka. A total of 164 cattle kidney samples were collected from the meat inspection hall in Colombo city during routine inspection procedures conducted by the municipal veterinary surgeons. The DNA was extracted and subjected to nested PCR for the detection of leptospiral flaB gene. Amplicons were sequenced, and phylogenic distances were calculated. Of 164 samples, 20 (12.2%) were positive for flaB‐PCR. Sequenced amplicons revealed that Leptospira species were deduced to L. borgpetersenii (10/20, 50%), L. kirschneri (7/20, 35%) and L. interrogans (3/20, 15%). The results indicate that a high proportion of the sampled cattle harbour a variety of pathogenic Leptospira spp, which can serve as important reservoirs for human disease.     

Similar frequency of Porcine circovirus 3 (PCV‐3) detection in serum samples of pigs affected by digestive or respiratory disorders and age‐matched clinically healthy pigs

Porcine circovirus 3 (PCV‐3) has been identified in pigs affected by different disease conditions, although its pathogenicity remains unclear. The objective of the present study was to assess the frequency of PCV‐3 infection in serum samples from animals suffering from post‐weaning respiratory or digestive disorders as well as in healthy animals. A total of 315 swine serum samples were analysed for PCV‐3 DNA detection by conventional PCR; positive samples were further assayed with a quantitative PCR and partially sequenced. Sera were obtained from 4 week‐ to 4 month‐old pigs clinically diagnosed with respiratory (n = 129) or digestive (n = 126) disorders. Serum samples of age‐matched healthy animals (n = 60) served as negative control. Pigs with clinical respiratory signs had a wide variety of pulmonary lesions including suppurative bronchopneumonia, interstitial pneumonia, fibrinous‐necrotizing pneumonia and/or pleuritis. Animals with enteric signs displayed histopathological findings like villus atrophy and fusion, catarrhal enteritis and/or catarrhal colitis. Overall, PCV‐3 DNA was detected in 19 out of 315 analysed samples (6.0%). Among the diseased animals, PCV‐3 was found in 6.2% (8 out of 129) and 5.6% (7 out of 126) of pigs with respiratory and digestive disorders, respectively. The frequency of PCV‐3 PCR positive samples among healthy pigs was 6.7% (4 out of 60). No apparent association was observed between PCR positive cases and any type of histopathological lesion. The phylogenetic analysis of the partial genome sequences obtained showed high identity among viruses from the three groups of animals studied. In conclusion, PCV‐3 was present in the serum of diseased and healthy pigs to similar percentages, suggesting that this virus does not seem to be causally associated with respiratory or enteric disorders.     

Immuno‐molecular prospecting for vector‐borne diseases in central Mexico

Climatic changes have influenced the temporal and spatial distribution of diseases. In livestock‐grazing areas, rodents are reservoirs of zoonotic pathogens; therefore, they play an important role in the transmission of diseases affecting domestic animals and humans. The objective of this study was to investigate the presence of the zoonotic agents: Anaplasma phagocytophilum, Borrelia burgdorferi, Ehrlichia canis and Rickettsia rickettsii, as well as the presence of viral RNA from the Bunyaviridae, Togaviridae and Flaviviridae families, in wild rodents from animal production units in central Mexico. The samples were obtained from wild rodents that had access and contact with animal production units. A total of 92 rodents were captured, and samples of blood, serum and organs, such as spleen, kidney, heart and liver, were obtained. The serum was used to detect antibodies against Anaplasma phagocytophilum, Borrelia burgdorferi, Ehrlichia canis and Rickettsia rickettsii by an immunofluorescence antibody test (IFAT); the blood was used for PCR analysis; and the organs were used to obtain RNA (cDNA) to perform RT‐PCR. By IFAT, all samples were positive to A. phagocytophilum and E. canis, and negative to B. burgdorferi and R. rickettsii. The samples that were positive to IFAT were used to confirm the presence of pathogen by PCR analysis. The results from the PCR were as follows: 34 samples were positive to A. phagocytophilum, and 59 to E. canis. There was no amplification of genetic material from the Bunyaviridae, Flaviviridae and Togaviridae virus families from the organs that were sampled, which suggests that the samples obtained did not contain RNA specific to these families. This is the first immuno‐molecular prospecting study on vector‐borne diseases in central Mexico demonstrating the presence of A. phagocytophilum and E. canis in wild rodents living in cattle grazing areas.     

Association d’une pneumopathie à Chlamydia psittaci et d’une maladie de Horton

Some arguments are in favor of the role of Chlamydia in the pathogenesis of atherosclerosis and some vasculitis. Illustrating this possible relation, we report the case of a patient developing consecutively a Chlamydia psittacci infection and a temporal arteritis. A 73-years-old woman, with no significant medical history, was hospitalized for constitutionnal symptoms. Three weeks before, she described fever and sore throat during 2 days. Since that, she remained exhausted and developped a mild intermittent limp of the jaws. Clinical examination was poor. A biological inflammatory syndrome was noticed. X-ray chest revealed bilateral interstitial opacities. The titer of anti-C. psittaci antibodies was significant (positive IgG at 1/ 2048). Soon after initiation of doxycyclin, a temporal arteritis biopsy performed, due to the persistance of clinical symptoms and high inflammatory syndrom, concluded to the diagnosis of temporal arteritis. Corticotherapy was added to antibiotherapy resulting in decreasing of inflammatory syndrom and improvement of general status of the patient. X-ray opacities decreased in 3 weeks. Serological control after 3 months showed a decrease of the titer of anti-C.psittacci antibodies to 1/256, confirming the initial diagnosis of Chlamydia pneumopathy. Our observation could constitute one more argument for the role of bacteria like Chlamydia in the pathogenesis of vascular diseases. Prospective seroepidemiologic and molecular biology studies could allow to clarify the association between Chlamydia infections and inflammatory vasculitis like temporal arteritis.     

Molecular detection of Leishmania spp. in Brazilian cross‐border south region mammalian hosts

Visceral leishmaniasis is an endemic zoonotic disease identified especially in developing territories. Brazil's northeast, southeast and midwest have been endemic for several years; currently, the infection is spreading to the south. Dogs are the main reservoirs; however, other mammal species have also been infected. Herein, we have identified the infecting Leishmania species in dogs and horses from the south of Brazil, a new outbreak of the infection. Blood samples were collected in the urban area of Uruguaiana city. DNA was extracted from peripheral blood, kinetoplast DNA (kDNA) and ribosomal DNA (rDNA) fragments were obtained by polymerase chain reaction (PCR) and sequenced. Out of 123 samples, 25 of them (14 dogs and 11 horses) were positive for Leishmania spp. Sequence alignment and phylogenetic analysis revealed that the kDNA in positive samples was similar to four species previously reported: L. infantum/L. chagasi, L. donovani, L. major. Despite kDNA minicircles regions are very useful due to high sensitivity to Leishmania spp. DNA detection, the sequence polymorphism among minicircles can be an obstacle to interspecific differentiation. Our results suggest that these strains are circulating in Brazil south region cross‐border and indicate the susceptibility of new outbreak for visceral leishmaniasis infection in horses domiciled in endemic region for canine and human visceral leishmaniasis.