Prevalence of Chlamydophila psittaci in Fecal Droppings from Feral Pigeons in Amsterdam, The Netherlands

In many cities, the feral rock dove is an abundant bird species that can harbor Chlamydophila psittaci. We determined the prevalence and genotype of C. psittaci in fresh fecal samples from feral pigeons in Amsterdam, The Netherlands. The prevalence was 7.9% overall (26/331; 95% confidence interval, 5 to 11). Ten genotyped PCR-positive samples were all genotype B.     

Genetic diversity and epizootiology of Chlamydophila psittaci prevalent among the captive and feral avian species based on VD2 region of ompA gene

To study genetic diversity and occurrence of Chlamydophila psittaci, a total of 1,147 samples from 11 avian orders including 53 genera and 113 species of feral and captive birds were examined using ompA gene based nested PCR. Three types of chlamydiae: C. psittaci (94.12%), C. abortus (4.41%) and unknown Chlamydophila sp. (1.47%) were identified among 68 (5.93%) positive samples (Psittaciformes-59, Ciconiiformes-8 and Passeriformes-1). Based on nucleotide sequence variations in the VD2 region of ompA gene, all 64 detected C. psittaci strains were grouped into 4 genetic clusters. Clusters I, II, III and IV were detected from 57.35%, 19.12%, 10.29% and 7.35% samples respectively. A single strain of unknown Chlamydophila sp. was found phylogenetically intermediate between Chlamydophila species infecting avian and mammalian hosts. Among Psittaciformes, 28 out of 81 tested species including 10 species previously unreported were found to be chlamydiae positive. Chlamydiosis was detected among 8.97% sick and 48.39% dead birds as well 4.43% clinically normal birds. Therefore, it was observed that though various genetically diverse chlamydiae may cause avian chlamydiosis, only a few C. psittaci strains are highly prevalent and frequently associated with clinical/subclinical infections.     

Monoclonal antibodies against Chlamydia psittaci

Five monoclonal antibodies were prepared against Chlamydia (C.) psittaci strain Pigeon-1041 isolated from a feral pigeon in Sapporo. Reactions of these antibodies to chlamydiae were examined using five strains of C. psittaci and two strains of C. trachomatis in an enzyme-linked immunosorbent assay, microimmunofluorescent test and complement fixation test. The antibodies were divided into two groups: three genus-specific (A2, D2, and I21) and two strain-specific (F2 and H9) antibodies. The antigenic determinant site of A2 was KIO4 sensitive, but those of D2, F2, and H9 were not affected greatly by KIO4 treatment. Nine C. psittaci strains from feral pigeons and 16 strains from budgerigars were classified into three groups and four groups, respectively, by reaction patterns against the monoclonal antibodies.     

三种方法对禽鹦鹉热嗜性衣原体疑似病例病原的流行调查

目的利用免疫荧光抗体法、PCE-ELISA、PCR法分别检测北京市及周边地区禽鹦鹉热嗜性衣原体疑似病例的病原,以了解和评价该病原流行状况。方法收集临床疑似禽衣原体的父母代、商品代家禽、SPF鸡喉头拭子、气囊组织、子宫黏膜,组织标本固定后采用直接免疫荧光染色测定衣原体包含体;喉头拭子、子宫黏膜处理后以PCE-ELISA定量测定衣原体;以衣原体保守性高的序列设计CTU/CTL引物扩增病料组织的omp-1基因片段。结果直接荧光染色法显示家禽衣原体平均阳性率为38.7%,组织检出率依次为子宫黏膜、气囊、喉头拭子;PCE-ELISA检测显示平均阳性率为58.7%,其中SPF鸡阳性率为10.0%,健康肉鸡达到30.0%;PCR法检测家禽衣原体平均阳性率为71.6%,SPF鸡为10.0%。样本的目的基因与标准禽衣原体6BC同源性超过99%。结论种禽场和商品养殖场均发现疑似病例中禽鹦鹉热嗜性衣原体感染情况较为严重。荧光抗体染色、PCE-ELISA可用于临床实践中进行快速、准确检测。     

Prevalence of infection and genetic diversity of porcine circovirus type 2 (PCV2) in wild boar (Sus scrofa) in Poland

Porcine circovirus type 2 (PCV2) is a widespread, important pathogen of domestic swine and the causative agent of postweaning multisystemic wasting syndrome and other diseases and conditions referred to as "porcine circovirus diseases." Specific antibodies and DNA to PCV2 have also been detected in European wild boars and North American feral pigs. We collected 312 tonsil samples from wild boars shot in 13 of 16 districts of Poland, and tested them for PCV2 DNA using a real-time PCR. We detected PCV2 DNA in 75.6% of tested tonsils, and in particular, in 60% of samples from the 2006-07 season, and 91% from 2007-08. The phylogenetic analysis that included 12 PCV2 sequences from wild boars revealed that they belonged to two genetic clusters, PCV2b and PCV2a. We present data on prevalence of PCV2 in Polish wild boars and for the first time report the PCV2a genotype in Poland.     

Prevalence of Toxoplasma gondii infection in feral cats in Seoul, Korea

The present study assessed the prevalence of Toxoplasma gondii infection in feral cat populations in Seoul using enzyme-linked immunosorbent assay (ELISA) and nested polymerase chain reaction (PCR). A total of 456 feral cats from 17 wards in Seoul was surveyed. The overall prevalence of T. gondii infection was 15.8% (69/456) by ELISA and 17.5% (80/456) by PCR; by gender, 17% (44/259) by ELISA and 16.2% (42/259) by PCR in males and 14.3% (28/196) by ELISA and 19.4% (38/196) by PCR in females. On a baseline of the Han River, prevalence was 15.1% (29/192) by ELISA and 15.6% (30/192) by PCR in the upper region and 16.4% (43/264) by ELISA and 18.9% (50/264) by PCR in the lower area. This suggested that toxoplasmosis is widespread throughout Seoul's feral cat population and it is critical that the city institute policies for the control of the feral cat population to reduce the risk of toxoplasmosis transmission to animals, including humans.     

Aspects of the epidemiology of Chlamydia psittaci infection in a population of koalas (Phascolarctos cinereus) in southeastern Queensland, Australia

A population of free-ranging koalas in southeastern Queensland was examined to determine the prevalence of Chlamydia psittaci infections. Although C. psittaci was isolated from 46 of 65 (71%) koalas studied, only six (9%) of these had clinical signs of disease. Most adult females (82%) had back or pouch young present even though 67% of them were infected. There were no significant correlations between age, sex or site of sampling (urogenital versus conjunctival tissues) and the isolation of C. psittaci. No other important bacterial or fungal pathogens were isolated. The complement fixation test had a sensitivity of 7% and a specificity of 94% in detecting chlamydial infections, suggesting that it is unsuitable for use as a screening test. Chlamydia psittaci infection within this population appeared to represent a generally well-balanced host-parasite relationship and few animals had clinical signs of disease. Only four of 27 (15%) healthy koalas infected with C. psittaci followed for 24 wk after sampling developed eye disease or "dirty tail." Two koalas with keratoconjunctivitis recovered without treatment during the study period. Additional factors, including the stresses imposed by loss of habitat, may act to produce overt disease in koalas with latent C. psittaci infections.     

Detection of Clostridium difficile and Salmonella in feral swine population in North Carolina

We sampled 161 feral pigs in eastern North Carolina, USA, to determine the prevalence and antimicrobial resistance profile of Clostridium difficile and Salmonella. Seven (4.4%) and eight (5.0%) pigs tested positive for C. difficile and Salmonella, respectively, highlighting the importance of determining the epidemiology of these pathogens in feral pigs.     

Newcastle disease virus and Chlamydia psittaci in free-living raptors from eastern Germany

Organ samples from free-living raptors from the federal states of Berlin and Brandenburg in eastern Germany were tested for Newcastle disease virus (NDV; n = 331) and Chlamydia psittaci (n = 39) by polymerase chain reaction (PCR). In 18 individuals NDV nucleic acids were detected. These samples originated from barn owls (Tyto alba; n = 15, 28%), tawny owl (Strix aluco; n = 1, 5%), common buzzard (Buteo buteo, n = 1, 1%), and European kestrel (Falco tinnunculus; n = 1, 4%). In 29 (74%) of 39 samples C. psittaci was detected. Chlamydia psittaci is common in free-living birds of prey in the investigated area.     

Detection of Streptobacillus spp. in feral rats by specific polymerase chain reaction

Streptobacillus moniliformis is an etiological agent of rat-bite fever and Haverhill fever in human infection. As the currently available methods for identifying the causative bacteria are not satisfactory, we attempted to establish them by PCR using newly designed primers for the 16S rRNA gene of S. moniliformis. We then determined the prevalence of Streptobacillus spp. in two species of feral rats that inhabit an urban region in Japan, because information on the prevalence of the bacteria in feral rats is obscure. The use of PCR with newly designed primers showed that an extremely high proportion of R. norvegicus harbored the bacteria (61/66, 92%), whereas the prevalence was only 58% in R. rattus (30/52). The nucleotide sequence analysis of the 16S rRNA gene of Streptobacillus spp. isolated from oral swabs of feral rats showed at least two different types of bacteria among isolates from R. norvegicus and R. rattus.     

Prevalence of and associated risk factors for shedding Cryptosporidium parvum oocysts and Giardia cysts within feral pig populations in California.

Populations of feral pigs (Sus scrofa) may serve as an environmental reservoir of Cryptosporidium parvum oocysts and Giardia sp. cysts for source water. We conducted a cross-sectional study to determine the prevalence of and associated demographic and environmental risk factors for the shedding of C. parvum oocysts and Giardia sp. cysts. Feral pigs were either live-trapped or dispatched from 10 populations located along the coastal mountains of western California, and fecal samples were obtained for immunofluorescence detection of C. parvum oocysts and Giardia sp. cysts. We found that 12 (5.4%) and 17 (7.6%) of 221 feral pigs were shedding C. parvum oocysts and Giardia sp. cysts, respectively. The pig's sex and body condition and the presence of cattle were not associated with the probability of the shedding of C. parvum oocysts. However, younger pigs (< or = 8 months) and pigs from high-density populations (> 2.0 feral pigs/km2) were significantly more likely to shed oocysts compared to older pigs (> 8 months) and pigs from low-density populations (< or = 1.9 feral pigs/km2). In contrast, none of these demographic and environmental variables were associated with the probability of the shedding of Giardia sp. cysts among feral pigs. These results suggest that given the propensity for feral pigs to focus their activity in riparian areas, feral pigs may serve as a source of protozoal contamination for surface water.     

Molecular genetic characterization of avian Chlamydophila psittaci isolates

Restriction enzyme analysis of the omp1 gene was used to characterize 51 avian Chlamydophila psittaci isolates. The analysis confirmed the predominance of genotype A in parrot C. psittaci isolates and revealed new omp1 genotypes in corvid C. psittaci isolates. The corvid isolates proved to lack an extrachromosomal plasmid. The omp1 and rRNA IGS sequences were determined for the new isolates. Phylogenetic analysis showed that isolate 1V, obtained from a crow, is intermediate in several characters between C. abortus and C. psittaci. The results were compared with data on the phylogenetic relationships of earlier chlamydium isolates.     

Quantitative detection of Porphyromonas gingivalis fimA genotypes in dental plaque

We developed quantitative fimA genotype assays and applied them in a pilot study investigating the fimbrial genotype distribution of Porphyromonas gingivalis in European subjects with or without chronic periodontitis. P. gingivalis was found in 71% and 9% of the samples from patients and healthy subjects, respectively. Enumeration of total P. gingivalis cell numbers by polymerase chain reaction and immunofluorescence showed excellent correspondence (r = 0.964). 73% of positive samples contained multiple fimA genotypes, but generally one genotype predominated by one to three orders of magnitude. Genotype II predominated in 60% of the samples. Genotype IV occurred with similar prevalence (73%) as genotype II but predominated in only 20% of the samples. Genotypes I, III and V were of much lower prevalence and cell densities of the latter two remained sparse. Our results suggest marked differences among the fimA genotypes' ability to colonize host sites with high cell numbers.     

Mesostigmatid mite infestations of rodents in diverse biotopes of central and southern India

Examination of 165 rodents trapped in domestic, peridomestic, and feral biotopes of central and southern India revealed the presence of 1,359 mesostigmatid mites. Rodents in central India were infested with 1 species of mite, Laelaps nuttalli; 98% of these mites were recovered from the peridomestic rodent Bandicota bengalensis. Material from the burrow nest of a peridomestic rodent examined in central India revealed the mite, Hypoaspis miles. Rodents from the diverse biotopes of southern India were infested by 8 species of mites: L. nuttalli, Laelaps buxtoni, Laelaps myonyssognathus, Androlaelaps aduncus, Androlaelaps marshalli, Androlaelaps sp. A, Hypoaspis sp. 1, and Hypoaspis sp. 2. Sheep from nearby villages regularly graze in feral biotopes. From the hair of these sheep, 2 species of mesostigmatid mites were recovered, A. aduncus and A. marshalli. The sheep appeared to aid the dispersal of rodent-infesting mites passively. Laelaps nuttalli was the predominant mite species found on the peridomestic rodent B. bengalensis. In both central and southern India, the prevalence, mean intensity, and relative density of this mite were much higher on peridomestic rodents than on domestic and feral rodents. A significant negative correlation was found between the numbers of mites and fleas infesting rodents in central India. A highly significant positive association between A. marshalli and A. aduncus on the feral rodent Tatera indica was recorded. In southern India, the overall prevalence of the 8 species of mites was highest on Mus platythrix. However, the combined prevalence of these mites on 2 feral rodents T. indica and M. platythrix was lower than their prevalence on B. bengalensis. Similarly, the combined values for mean intensity and relative density of these mites on the 2 feral rodents were lower than on the peridomestic rodent B. bengalensis.     

Absence of evidence is not evidence of absence: Feral pigs as vectors of soil‐borne pathogens

Invasive soil‐borne pathogens are a major threat to forest ecosystems worldwide. The newly discovered soil pathogen, Phytophthora ‘taxon Agathis’ (PTA), is a serious threat to endemic kauri (Agathis australis: Araucariaceae) in New Zealand. This study examined the potential for feral pigs to act as vectors of PTA. We investigated whether snouts and trotters of feral pigs carry soil contaminated with PTA, and using these results determined the probability that feral pigs act as a vector. We screened the soil on trotters and snouts from 457 pigs for PTA using various baiting techniques and molecular testing. This study detected 19 species of plant pathogens in the soil on pig trotters and snouts, including a different Phytophthora species (Phytophthora cinnamomi). However, no PTA was isolated from the samples. A positive control experiment showed a test sensitivity of 0–3% for the baiting methods and the data obtained were used in a Bayesian probability modelling approach. This showed a posterior probability of 35–90% (dependent on test sensitivity scores and design prevalence) that pigs do vector PTA and estimated that a sample size of over 1000 trotters would be required to prove a negative result. We conclude that feral pigs cannot be ruled out as a vector of soil‐based plant pathogens and that there is still a high probability that feral pigs do vector PTA, despite our negative results. We also highlight the need to develop a more sensitive test for PTA in small soil samples associated with pigs due to unreliable detection rates using the current method.     

Prevalence of seropositivity to pathogens in small carnivores in adjacent areas of Lazovskii Reserve

The prevalence of infectious diseases in wild and feral carnivores is poorly known in Primorsky Krai, where rare species such as the Amur tiger and the Far East leopard roam. In this study we evaluated the prevalence of seropositivity in feral (cats, dogs) and wild (raccoon dog, badger, American mink, Far Eastern wild cat) carnivores to various pathogens: distemper virus, parvovirus, feline immunodeficiency and feline leukemia virus, feline coronavirus, Toxoplasma gondii, Mycoplasma sp., and Chlamydia sp. It was shown that seropositivity occurred significantly more frequent in feral animals than in wild ones. The highest percentage of seropositive animals was observed in feral dogs. It is these diseases that can be dangerous for the Amur tiger and the Far East leopard, thus affecting their populations.     

Prevalence and transmission of pseudorabies virus in an isolated population of feral swine

Six hundred sixty-one feral swine (Sus scrofa) from Ossabaw Island, Georgia (USA) were captured, bled, and their sera tested for pseudorabies virus (PRV) antibody during a 6 yr period. Prevalence of seroconversion in females was somewhat higher than in males (10% versus 7%), but the difference was not statistically significant. Adults had a significantly higher prevalence than juveniles (29% versus 1%). An important finding in this study was that seroconversion occurred primarily in the adult feral swine.     

Prevalence, genotypes and risk factors associated with hepatitis C virus infection in hemodialysis patients in Campo Grande, MS, Brazil

A survey was conducted among the hemodialysis units of the city of Campo Grande, located in the state of Mato Grosso do Sul in the Mid-west region of Brazil, with the aim of investigating the prevalence, risk factors, and genotypes of hepatitis C virus (HCV) infection. A total of 163 patients were interviewed in five dialysis units. Serum samples were screened for anti-HCV. Positive samples were tested for HCV RNA and genotyped. The prevalence of anti-HCV was 11% (95% CI: 6.8-17.1). A history of transfusion with blood that was not screened for anti-HCV and length of time on hemodialysis were associated with HCV infection. HCV RNA was detected in 12 samples: ten were of genotype 1, subtypes 1a (75%) and 1b (8.3%), and two were of genotype 3, subtype 3a (16.7%).     

GB virus C/hepatitis G virus infection in dialysis patients and kidney transplant recipients in Central Brazil

In order to investigate the prevalence of GB virus C (GBV-C)/hepatitis G virus (HGV) infection in dialysis patients and kidney transplant recipients in Central Brazil and also to analyze the virus genotypes distribution, a total of 123 patients including 98 on hemodialysis, 13 on continuous ambulatory peritoneal dialysis treatment, and 12 who received kidney transplantation were interviewed in one unit of dialysis treatment in Goiania city. Blood samples were collected and serum samples tested for GBV-C/HGV RNA by polymerase chain reaction. Genotypes were determined by restriction fragment length polymorphism (RFLP) analysis. Eighteen samples were GBV-C/HGV RNA-positive, resulting in an overall prevalence of 14.6% (95% CI: 9.2-21.7). A high positivity for GBV-C/HGV RNA was observed in patients who had received kidney transplant (16.7%), followed by those on hemodialysis (15.3%), and peritoneal dialysis (7.7%). RFLP analysis revealed the presence of genotypes 1, 2, and 3 of GBV-C/HGV; more precisely, 9 (50%) samples were found belonging to the 2b subtype, 4 (22%) to the 2a subtype, 3 (17%) to genotype 1, and 2 (11%) to genotype 3. The present data indicate an intermediate prevalence of GBV-C/HGV infection among dialysis patients and kidney transplant recipients in Central Brazil. Genotype 2 (subtype 2b) seems to be the most prevalent GBV-C/HGV genotype in our region.     

Occurrence of Transgenic Feral Alfalfa (Medicago sativa subsp. sativa L.) in Alfalfa Seed Production Areas in the United States

The potential environmental risks of transgene exposure are not clear for alfalfa (Medicago sativa subsp. sativa), a perennial crop that is cross-pollinated by insects. We gathered data on feral alfalfa in major alfalfa seed-production areas in the western United States to (1) evaluate evidence that feral transgenic plants spread transgenes and (2) determine environmental and agricultural production factors influencing the location of feral alfalfa, especially transgenic plants. Road verges in Fresno, California; Canyon, Idaho; and Walla Walla, Washington were surveyed in 2011 and 2012 for feral plants, and samples were tested for the CP4 EPSPS protein that conveys resistance to glyphosate. Of 4580 sites surveyed, feral plants were observed at 404 sites. Twenty-seven percent of these sites had transgenic plants. The frequency of sites having transgenic feral plants varied among our study areas. Transgenic plants were found in 32.7%, 21.4.7% and 8.3% of feral plant sites in Fresno, Canyon and Walla Walla, respectively. Spatial analysis suggested that feral populations started independently and tended to cluster in seed and hay production areas, places where seed tended to drop. Significant but low spatial auto correlation suggested that in some instances, plants colonized nearby locations. Neighboring feral plants were frequently within pollinator foraging range; however, further research is needed to confirm transgene flow. Locations of feral plant clusters were not well predicted by environmental and production variables. However, the likelihood of seed spillage during production and transport had predictive value in explaining the occurrence of transgenic feral populations. Our study confirms that genetically engineered alfalfa has dispersed into the environment, and suggests that minimizing seed spillage and eradicating feral alfalfa along road sides would be effective strategies to minimize transgene dispersal.