Antimicrobial Sensitivity Profile in Psittacine Birds at an Avian Teaching Hospital: A Retrospective Study, 2015-2022.

Veterinary hospitals house patient populations with diverse infectious statuses, microbiota, and histories of prior antibiotic therapy. Choanal swabs are commonly used for assessing the upper respiratory tract of birds for bacterial disease, with the samples submitted for cytologic testing and/or culture and antimicrobial sensitivity testing. The aim of this retrospective study was to identify and quantify bacteria isolated from choanal swabs collected from psittacine patients at a veterinary teaching hospital in Mexico City, Mexico. Data regarding bacterial isolates from choanal swabs were obtained from the medical records of companion psittacines suspected of upper respiratory bacterial disease that presented between November 2015 and December 2022. A total of 47.8% (175 of 366) of the bacterial isolates were from specimens obtained from red-lored Amazons (Amazona autumnalis). Gram-negative bacteria predominated, with 27 different genera identified. Klebsiella, Staphylococcus, and Escherichia were the most frequently isolated genera. A total of 90.4% (331 of 366) of the isolates were resistant to at least 1 antibiotic tested in the sensitivity panel, and a single Klebsiella isolate was resistant to 13 different antibiotics. Gentamicin had a high percentage of efficacy (79.5%; 182 of 229) against the bacterial isolates, whereas isolates tested against sulfonamide-trimethoprim (46.7%, 98 of 210), streptomycin (43.8%; 88 of 201), and clindamycin (12.9%; 15 of 116) had susceptibilities <50%. This is the first study to report common bacterial isolates and their antimicrobial susceptibility patterns from choanal swab samples collected from companion psittacines suspected of upper respiratory disease in Mexico. Clinicians can use the information presented in this study as a guide for therapeutic decision-making when managing upper respiratory bacterial infections in companion psittacine patients.

Ciprofloxacin enhances therapeutic levels of voriconazole through CYP450 inhibition in the common raven (Corvus corax), possibly improving efficacy against aspergillosis: a pilot study.

OBJECTIVE: To determine if a cytochrome (CYP) P450 enzyme inhibitor can maintain therapeutic plasma levels of voriconazole when administered orally. ANIMALS: 11 healthy, common ravens (Corvus corax). METHODS: Birds were randomly assigned to pilot study groups to receive voriconazole orally alone or combined with a CYP inhibitor. Pilot studies with 3 CYP inhibitors launched the main study using ciprofloxacin (20 mg/kg) followed 1 hour later by voriconazole (6 mg/kg) every 12 hours for 14 days. Plasma voriconazole concentrations were measured at various time points by HPLC-MS. The study period lasted from September 2016 to December 2020. RESULTS: The birds failed to maintain therapeutic plasma levels of voriconazole during multidose administration alone or following preadministration with various CYP inhibitors. For the 14-day study period, voriconazole reached a maximum plasma concentration of 2.99 µg/mL with a time-to-peak drug concentration of 1.2 hours following preadministration of ciprofloxacin. One bird was removed from the study due to lethargy, but the other birds completed the study without incident. CLINICAL RELEVANCE: Ciprofloxacin (20 mg/kg) followed by voriconazole (6 mg/kg) maintained the concentration of voriconazole within the recommended therapeutic range of 0.5 to 5 µg/mL without toxicity. Ciprofloxacin prevented the saturable metabolism of voriconazole and maintained these levels for the study duration. This drug combination could be used in the treatment of chronic aspergillosis in the common raven.

Pathological findings and their association with diseases of captive psittacine birds native to Mexico.

The captivity and use of native psittacine birds is prohibited in Mexico. However, as these birds are among the groups most affected by illegal trafficking, they are commonly found as companion animals. Nevertheless, it is difficult to obtain information on their health. Therefore, a retrospective study was conducted of the clinical histories and necropsy reports of native psittacines that had been submitted to the Bird Disease Diagnostic and Research Laboratory of the Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, from 2006 to 2017. The lesions were classified according to type and anatomical location and the diseases were classified as infectious or non-infectious. During this period, 252 psittacines were submitted, the most frequent of which were the red-lored parrot (Amazona autumnalis), orange-fronted parakeet (Eupsittula canicularis) and scarlet macaw (Ara macao). The lesions were primarily located in the digestive and respiratory systems. By integrating the clinical histories and post-mortem findings, we concluded that nutritional disorders were the most frequent non-infectious diseases, systemic bacterial infections were the most frequent infectious conditions, the primary parasite was Sarcocystis spp and the most frequent neoplasm was multicentric lymphoma.

An Acute Mortality Event Associated with Novel Macrorhabdus ornithogaster Infection and Underlying Factors in a Newly-Established Captive Group of American White Ibis (Eudocimus albus) Nestlings.

Twenty-four American white ibis (Eudocimus albus) nestlings were collected in Florida (USA) on 17 April 2017 to establish a captive flock. On 7 May 2017, three birds died suddenly, following severe lethargy, hemorrhaging from the mouth and nares, anorexia, and production of bright-green colored feces. An additional ibis with delayed growth and pathological fractures was euthanized 18 May 2017. Severe ventriculitis associated with Macrorhabdus ornithogaster was noted in all four birds, bacterial sepsis was confirmed in one bird by culture and histologic examination, and bacterial endotoxemia was suspected in two birds based on gross and histologic examination, but no bacteria were isolated from these birds. Birds also had vitamin E liver levels consistent with coagulopathy previously described in pelicans. We sampled feces from 91 adult, free-living, healthy ibis in Florida in July 2017 and found 71% were shedding organisms with morphologic characteristics consistent with Macrorhabdus sp. Molecular characterization of the ibis-origin M. ornithogaster showed it was phylogenetically related to numerous M. ornithogaster sequences. It is unknown if M. ornithogaster infection resulted in clinical disease as a result of dietary or stress-related dysbiosis, or other factors. Macrorhabdus-associated disease has not previously been confirmed in wading birds. We discuss potential associations of gastric M. ornithogaster infection with morbidity and mortality in these cases and highlight the need for additional studies on this pathogen in free-living birds.

Determining the Prevalence of Avian Chlamydiosis in Wild Amazona Species From Brazil Using Molecular Testing and Clinical Signs.

Avian chlamydiosis is a disease that occurs in birds, especially parrots, and is caused by the Gram-negative bacterium Chlamydia psittaci. Wild Animal Screening Centers in Brazil receive, maintain, treat, and place (preferably to nature) wild animals recovered from illegal trafficking. We performed molecular testing for avian chlamydiosis in parrots from the genus Amazona that were presented to these centers. Cloacal swab samples were collected from 59 parrots (Amazona species) and transported in aqueous or culture medium. The samples were subsequently submitted for DNA extraction by the boiling method, polymerase chain reaction (PCR) amplification using CPF/CPR primers, and agarose gel electrophoresis. Conjunctivitis, nasal discharge, and poor body condition were the clinical signs associated with a differential disease diagnosis of avian chlamydiosis. Transport medium did not have an effect on the test results. The prevalence of C psittaci in the samples was 37% (22/59, 95% confidence interval: 25-49). There was a significant (P = 0.009) association between the PCR test results and clinical signs. Follow-up testing was conducted on a subgroup of 14 individuals that initially tested negative on PCR; 50% (7/14) of these birds were found to be positive within 24 days of the first test. The results of this study confirm the feasibility of using the CPF/CFP primer-based PCR to detect C psittaci in Amazona species, describe a less costly method of transporting biological material for DNA extraction, and evaluate the temporal aspect for obtaining positive results through molecular testing for C psittaci in Amazona species.

Application of environmental sampling to investigate a case of avian chlamydiosis in a pet store and breeding facility leading to mass bird exposures.

Chlamydia psittaci is a bacterium that causes chlamydiosis in birds and can cause zoonotic psittacosis in people. In November 2017, we received notification of a suspected case of avian chlamydiosis in a captive cockatiel (Nymphicus hollandicus) that was sold by an online pet bird retail and breeding facility in Washington State. We describe the investigation with emphasis on how environmental sampling was used to guide veterinary and public health interventions. Bird samples were collected either from pooled droppings, pooled plumage or individual nasal and choanal swabs. Environmental samples were obtained by swabbing cleaning mops, tables and cage structures. All samples were tested by polymerase chain reaction and positive samples underwent genotyping. Approximately 1000 birds representing four taxonomic orders were kept within an open-space warehouse. Eight of 14 environmental samples and one of two pooled faecal samples were positive for Chlamydia spp. The contaminating strain of Chlamydia spp. was identified as genotype A. The facility was closed for environmental disinfection, and all psittacines were treated with oral doxycycline for 45 days. Ten of 10 environmental and two of two pooled faecal samples were negative for C. psittaci 11 months after the completion of environmental disinfection and antimicrobial treatment. This investigation highlights the importance of preventing and mitigating pathogen incursion in an online pet retail and breeding facility. Environmental sampling is valuable to guide animal and public health interventions for control of C. psittaci, particularly when large numbers of birds are exposed to the pathogen.

Are Purple Finches (Haemorhous purpureus) the Next Host for a Mycoplasmal Conjunctivitis Epidemic?

Ever since 1994, when the bacterial pathogen Mycoplasma gallisepticum jumped from poultry to wild birds, it has been assumed that the primary host species of this pathogen in wild North American birds was the house finch (Haemorhous mexicanus), in which disease prevalence was higher than in any other bird species. Here we tested two hypotheses to explain a recent increase in disease prevalence in purple finches (Haemorhous purpureus) around Ithaca, New York. Hypothesis 1 is that, as M. gallisepticum evolved and became more virulent, it has also become better adapted to other finches. If this is correct, early isolates of M. gallisepticum should cause less-severe eye lesions in purple finches than in house finches, while more-recent isolates should cause eye lesions of similar severity in the two species. Hypothesis 2 is that, as house finch abundance declined following the M. gallisepticum epidemic, purple finches around Ithaca increased in abundance relative to house finches and purple finches are thus more frequently exposed to M. gallisepticum-infected house finches. This would then lead to an increase in M. gallisepticum prevalence in purple finches. Following an experimental infection with an early and a more-recent M. gallisepticum isolate, eye lesions in purple finches were more severe than in house finches. This did not a support Hypothesis 1; similarly, an analysis of Project Feeder Watch data collected around Ithaca did not show differences in changes in purple and house finches' abundance since 2006, a result which does not support Hypothesis 2. We conclude that purple finch populations will, unlike those of house finches, not suffer a severe decline because of a M. gallisepticum epidemic.

¿Son los pinzones purpúreos (Haemorhous purpureus) los próximos huéspedes de una epidemia de conjuntivitis por micoplasma? Desde el año 1994, cuando el patógeno bacteriano Mycoplasma gallisepticum saltó de las aves comerciales a las aves silvestres, se ha supuesto que la principal especie huésped de este patógeno en las aves silvestres de América del Norte era el pinzón mexicano (Haemorhous mexicanus), en el que la prevalencia de la enfermedad era mayor que en cualquier otra especie aviar. En este estudio se analizaron dos hipótesis para explicar un aumento reciente en la prevalencia de la enfermedad en los pinzones purpúreos (Haemorhous purpureus) alrededor de Ithaca, en Nueva York. La hipótesis 1 es que, a medida que M. gallisepticum evolucionó y se volvió más virulento, también se adaptó mejor a otros pinzones. Si esto es correcto, los aislamientos tempranos de M. gallisepticum deberían causar lesiones oculares menos graves en los pinzones purpúreos que en los pinzones mexicanos, mientras que los aislamientos más recientes deberían causar lesiones oculares de gravedad similar en las dos especies. La hipótesis 2 es que, a medida que la abundancia de pinzones mexicanos disminuyó después de la epidemia de M. gallisepticum, los pinzones purpúreos alrededor de Ithaca aumentaron en abundancia en relación con los pinzones mexicanos y, por lo tanto, los pinzones morados están expuestos con mayor frecuencia a los pinzones caseros infectados con M. gallisepticum. Esto conduciría a un aumento de la prevalencia de M. gallisepticum en los pinzones purpúreos. Después de una infección experimental con un aislamiento temprano y uno más reciente de M. gallisepticum, las lesiones oculares en los pinzones purpúreos fueron más graves que en los pinzones mexicanos. Esto no apoyó la Hipótesis 1; de manera similar, un análisis de los datos del Proyecto Feeder Watch recopilados alrededor de Ithaca no mostró diferencias en los cambios de la abundancia de pinzones purpúreos y mexicanos desde 2006, un resultado que no respalda la Hipótesis 2. Se concluye que las poblaciones de pinzones purpúreos, a diferencia de las de los pinzones mexicanos, no sufrieron un declive severo a causa de una epidemia de M. gallisepticum.

Prevalence of asymptomatic infections of Chlamydia psittaci in psittacine birds in Korea.

Avian chlamydiosis is an acute or chronic bacterial disease of birds. Chlamydia psittaci is the primary agent of the disease. It is also an important zoonotic pathogen. Chlamydia avium and Chlamydia gallinacea have also been recognized as potential causative agents of the disease. Clinical signs of this disease can vary in severity. Asymptomatic infections of Chlamydia have commonly been reported in various birds worldwide. In this study, we investigated the distribution of Chlamydia species in healthy psittacine birds in Korea. A total of 263 samples (pharyngeal/cloacal swabs and faeces) were collected from psittacine birds of 26 species in five zoos, five parrot farms and seven parrot cafes between 2020 and 2021. Ages of these birds had a wide range (1 month to 30 years). During sample collection, no bird showed any clinical signs indicating diseases such as chlamydiosis. Samples were tested for the presence of Chlamydia spp. using real-time PCR assays. Chlamydia spp. were detected in 168 (63.9%) samples and C. psittaci was detected in 96 (36.5%) samples. However, C. avium and C. gallinacea were not detected. There were no significant differences in the prevalence of asymptomatic infections in birds among three types of housing environments. Regarding ompA genotypes, 87 C. psittaci-positive samples had genotype A based on sequence analysis (n = 28) and genotype-specific real-time PCR (n = 59). Other positive samples were untyped (n = 9). Overall findings showed high prevalence of asymptomatic infections of C. psittaci in psittacine birds in Korea, posing a significant hazard to public health.

Immature hard ticks infected with Rickettsia amblyommatis on breeding birds from Pantanal.

Immature hard ticks from the genus Amblyomma feed on blood from a wide range of Neotropical avian hosts. They serve as vectors for pathogens of medical and veterinary importance, such as Rickettsia agents of the spotted fever group (SFG). Hence, determining ecological factors that increase encounter rates between immature ticks and their avian hosts may contribute to the understanding of tick-borne diseases transmission. Here, we used 720 individual birds from 96 species surveyed in the Brazilian Pantanal to test whether host breeding season influenced tick infestation probabilities. Additionally, collected ticks were screened for Rickettsia agents to describe new avian-tick-bacteria associations. Our models revealed that the probability of an individual bird being infested with immature ticks was similar during the breeding and pre-breeding season, but higher loads of immature tick stages were found during the breeding season. Host sex did not predict infestation probability, but Rickettsia agents recovered from ticks were more prevalent during the pre-breeding season. The new records of host usage by larvae and nymphs of Amblyomma in Pantanal and the growing body of tick surveys in Neotropical avian communities, suggest that immature ticks may benefit from avian blood sources during their annual cycle. The low number of infected ticks with Rickettsia agents on Pantanal birds suggest that this vertebrate group are likely not acting as reservoirs for these microorganisms. However, long-term surveys at the same site are imperative to determine which tick species are acting as reservoirs for Rickettsia agents in Pantanal and determine whether birds are playing a role in dispersing ticks and tick-borne pathogens.

Detection of Salmonella spp. in wild and domestic birds in an anthropized ecotone between the Cerrado and the Amazon Forest in Brazil.

Emergence of zoonotic infectious diseases represent one of the main threats to people worldwide. To properly understand and prevent zoonoses is fundamental to study their epidemiology and the possibility of spillover events, especially for commercially intensive domestic animals and humans. Here, we studied 210 wild birds from the "Ipucas" region, which consists of fragments of the Amazon Forest interspersed with fragments of the "Cerrado" that is subject to seasonal flooding and 75 domestic birds from neighboring poultry farming. Then, we molecularly diagnosed Salmonella and Chlamydia from wild birds and poultry. Among the wild birds, four were diagnosed with Chlamydia psittaci and 23 with Salmonella spp., while we detected 15 poultry infected by Salmonella spp. and no poultry with C. psittaci. We highlighted the common infections of wild and domestic birds in an anthropologically modified environment and potential spillover of Salmonella pathogens among wild and livestock birds. Those infections can harm the health of native and domestic species.

First detection of Chlamydia avium in healthy Amazon parrots (Amazona aestiva) in Argentina.

In recent years, new Chlamydia species, other than Chlamydia psittaci, have been confirmed in birds. One of these new species, Chlamydia avium, was reported mainly in pigeons and parrots in Europe. Analyzing multimucosal swabs obtained from 7 Amazon parrots (Amazona aestiva) from illegal trade and admitted to the Reserva Experimental Horco Molle (Tucuman, Argentina) for their rehabilitation, we describe the finding of the genetic material of C. avium in 2 of these birds. There were no signs compatible with the chlamydiosis-like disease in the studied birds or in the rehabilitation center staff. The use of sensitive and wide-ranging molecular tools is necessary for the detection of all Chlamydiaceae present in birds and would aid in the selection of control measures in wildlife rehabilitation centers to prevent outbreaks in the facilities and the introduction of pathogens in nature. We provide the first molecular evidence of the presence of C. avium in Argentina and a new species of psittacine host.

Causes of mortality of kiwi (Apteryx spp.) in New Zealand: a retrospective analysis of post-mortem records, 2010-2020.

AIMS: To examine and assess causes of mortality of kiwi (Apteryx spp.) submitted to Massey University between 2010 and 2020 across the five recognised species according to location, age group and captivity status in New Zealand. METHODS: Post-mortem reports were obtained from the Massey University/Te Kunenga ki Purehuroa School of Veterinary Science/Wildbase Pathology Register. Inclusion criteria were all species of kiwi with a date of post-mortem examination between August 2010 and August 2020. Data from each report was exported, categorised and compared using Microsoft Excel. RESULTS: Of a total of 1,005 post-mortem reports, there were 766 North Island brown kiwi (NIBK; A. mantelli), 83 tokoeka (A. australis), 73 rowi (A. rowi), 49 great spotted kiwi (A. haastii), and 34 little spotted kiwi (A. owenii). This comprised 19 eggs/embryos, 125 neonatal, 473 juvenile, 153 subadult, and 235 adult kiwi. There were 615 kiwi from wild populations, 148 from sanctuary populations, 238 from captivity, and four from unspecified locations. The leading cause of death was trauma, affecting 322 (32.0 (95% CI = 29.2-35.0)%) kiwi including 289 (37.3 (95% CI = 26.0-31.7)%) NIBK. Nearly half of these died from predation by mustelids, with losses recorded from neonates to adults and clustered in the central to southern North Island. Predation by dogs was the second most common cause of death, killing 84 (8.4 (95% CI = 6.7-10.2)%) kiwi, of which 65.5% came from the northern districts of the North Island. Non-infectious disease killed 214 (21 (95% CI = 18.8-24.0)%) kiwi, and included developmental deformities, gastrointestinal foreign bodies and predator trap injuries. Infectious disease killed 181 (18.0 (95% CI = 15.7-20.5)%) kiwi and the proportion decreased with age, with common diagnoses including coccidiosis, bacterial septicaemia, avian malaria, and fungal respiratory disease. Starvation affected 42 (4.2 (95% CI = 3.0-5.6)%) kiwi, comprised of mainly neonatal or juvenile individuals from wild or sanctuary populations, with a higher percentage seen in tokoeka (11/83; 13.3%) compared to other species (min 0%, max 5.9%). The cause of death was undetermined in 246 (24.5 (95% CI = 21.8-27.3)%) cases, which was most often due to poor preservation of remains. This included 33/73 (46%) rowi and 32/83 (39%) tokoeka, and affected mainly birds from sanctuary and wild populations. CONCLUSIONS: This study enhances our understanding of causes of mortality in captive, wild and sanctuary populations of all kiwi species and age groups within contemporary New Zealand.

Analysis for the diagnostic accuracy of PCR detection of Macrorhabdus in companion birds.

Background: Macrorhabdus ornithogaster, a yeast-like fungus, has the potential to infect various bird species, including companion birds. Although birds infected with M. ornithogaster may often remain asymptomatic, the infection can develop into chronic wasting gastritis and even progress to gastric cancer, highlighting the importance of early detection of M. ornithogaster infection. Despite direct fecal examination being a commonly used diagnostic method, the polymerase chain reaction (PCR) is anticipated to offer a higher detection rate. However, the actual diagnostic accuracy of the PCR for M. ornithogaster remains unknown. Case Description: Ninety fecal samples collected from companion birds that visited or were admitted to a hospital, regardless of their stage of Macrorhabdus diagnosis or treatment, were subjected to PCR testing. An accuracy analysis was then performed, considering symptomatology, direct fecal testing (FT), and sequencing. The PCR test had a sensitivity of 83.33%, specificity of 95.00%, false negative rate of 16.67%, false positive rate of 5.00%, positive predictive value of 89.29%, negative predictive value of 91.94%, prevalence of 33.33%, positive likelihood ratio of 16.67, negative likelihood ratio of 0.18, and diagnostic odds ratio of 95.00. Conclusion: The findings suggest that the PCR for Macrorhabdus possesses high diagnostic accuracy, with the ability to accurately identify uninfected individuals as negative. While the direct fecal examination is appropriate for routine primary screening, in cases where M. ornithogaster is not detected by FT, the PCR may provide a more accurate and definitive diagnosis due to its high specificity.

Survey of respiratory disease associated with Mycoplasma gallisepticum in British gamebirds (2016-2019): In vitro antibiotic sensitivity, pathology and detection of other pathogens.

BACKGROUND: The causes of respiratory disease in British gamebirds were investigated during 2016-2019 following concerns about poorer responses to antibiotic treatment. Emphasis was placed on Mycoplasma gallisepticum, but other possible bacterial and viral causes were included, along with gross and histopathological examination. METHODS: Clinical respiratory disease outbreaks were investigated. RESULTS: Mycoplasma gallisepticum was detected by PCR in 65 of 69 outbreaks in pheasants and partridges and isolated from 56 of these. Partial mgc2 gene sequences from 28 M. gallisepticum isolates were compared, and 26 proved identical, suggesting the prevalence of a dominant sequence type. Minimum inhibitory concentration values for tiamulin, tylosin, tylvalosin, doxycycline and tetracycline were significantly higher than the reference strain but could not be correlated with treatment failures. Other bacterial species were isolated from sinuses but were not consistently correlated with disease. RT-PCRs detected coronaviruses in 18% of 49 outbreaks and avian metapneumovirus in 8%. Histopathological lesions were typical of M. gallisepticum sinusitis and significantly associated with M. gallisepticum PCR outbreak positivity. CONCLUSION: Mycoplasma gallisepticum remains an important cause of respiratory disease in gamebirds. Synergism with other pathogens may have played a role in some outbreaks. Specific reasons for variable responses to antibacterial treatment were not identified.

Ixodid diversity and detection of spotted fever group Rickettsia spp. in ticks collected on birds in the Brazilian Atlantic Forest.

The Brazilian Atlantic Forest helds one of the most diverse and unique avifauna in the world. Many vertebrate species are reservoirs of tick-borne pathogens, and birds are an important group among them due to their mobility which facilitates the dispersion of ticks and the infectious agents they carry. This study brings data on the tick diversity parasitizing birds and the molecular detection of Rickettsia spp. in these arthropods. Birds (n = 773) were captured, identified, and banded at Mata do Paraíso Research, Training, and Environmental Education Center located in Viçosa, Minas Gerais, Brazil. Birds were checked for the presence of ticks, which were individually collected, identified, and molecularly processed through Polymerase Chain Reaction (PCR) for the detection of Rickettsia spp. A total of 130 individuals were infested by ticks, and 479 tick specimens were collected, showing a seasonal distribution of the life stages throughout the year. Ticks were identified as Amblyomma longirostre (59/479); Amblyomma calcaratum (20/479); Amblyomma varium (3/479); Amblyomma sculptum (2/479) and Amblyomma spp. larvae (395/479). Seasonal distribution of the life stages of ticks was observed along the year and significant negative correlations were found between temperature and collected ticks and temperature and infested birds. From the evaluated samples of ticks, 25.44% (n = 43/169) scored positive for Rickettsia spp., and sequence analysis indicated high nucleotide identity with Rickettsia rhipicephali, R. massiliae, R. africae and R. honei marmionii. The potential for dispersal of ticks by birds added to the aggressiveness of species of the genus Amblyomma and the zoonotic potential of some species of Rickettsia are quite worrying when we consider that the study area is widely attended by students, researchers, people from the city and neighboring municipalities.

Genome Engineering in Mycoplasma gallisepticum Using Exogenous Recombination Systems.

Mycoplasma gallisepticum (Mgal) is a common pathogen of poultry worldwide that has recently spread to North American house finches after a single host shift in 1994. The molecular determinants of Mgal virulence and host specificity are still largely unknown, mostly due to the absence of efficient methods for functional genomics. After evaluating two exogenous recombination systems derived from phages found in the phylogenetically related Spiroplasma phoeniceum and the more distant Bacillus subtilis, the RecET-like system from B. subtilis was successfully used for gene inactivation and targeted replacement in Mgal. In a second step, the Cre-lox recombination system was used for the removal of the antibiotic resistance marker in recombinant mutants. This study therefore describes the first genetic tool for targeted genome engineering of Mgal and demonstrates the efficiency of heterologous recombination systems in minimal bacteria.

The prevalence of Chlamydia psittaci in confiscated Psittacidae in Colombia.

Chlamydia psittaci is a highly zoonotic bacteria distributed worldwide; it is responsible for psittacosis, one of the most important infectious diseases affecting the Psittacidae, mostly parrots. This work was aimed at determining C. psittaci prevalence and genotype in 177 parrots confiscated in Colombia; cloacal swab (166) and faecal (177) samples were analysed from birds confiscated and housed in a Temporary Wildlife Reception Centre (Centro de Reception de Fauna Temporal). Conventional PCR was run on the samples for amplifying the MOMP gene and then the ompA gene. The C. psittaci genotype A was found in 81.3 % (144/177) of the birds analysed. Cloacal swabs accounted for 129/166 (77.7 %) positive samples and faecal matter for 53/177 (29.9 %), 38 birds proving positive for both types of sample; there was an 8.15 times greater probability of detection for cloacal swabs compared to faecal swabs (p < 0.05). Clinical examination findings were correlated with the animals' positivity for cloacal swabs, faecal matter or both, finding a statistically significant relationship with low respiratory rate (p < 0.05) and broken plumage for cloacal swab sample results (p < 0.1). Even though 85 % seroprevalence has previously been reported in Colombia using indirect ELISA, this study reports for the first time C. psittaci genotype A endemicity in psittacines in captivity in Colombia using molecular techniques, considering the zoonotic risk involved in having these birds as pets.

A multiplex real-time PCR assay for differential identification of avian Chlamydia.

Avian chlamydiosis is an acute or chronic disease of birds after infection by Chlamydia. Although Chlamydia psittaci is the primary agent of the disease, two additional species, Chlamydia avium and Chlamydia gallinacea, have also been recognized as potential disease agents. Therefore, the diagnosis of avian chlamydiosis requires differential identification of these avian Chlamydia species. The objective of the present study was to develop a multiplex real-time polymerase chain reaction (PCR) assay to rapidly differentiate between these three species of avian Chlamydia (C. psittaci, C. avium, and C. gallinacea) as well as to detect the genus Chlamydia. Specific genetic regions of the three species were identified by comparative analysis of their genome sequences. Also, the genus-specific region was selected based on 23S rRNA sequences. PCR primers and probes specific to the genus and each species were designed and integrated in the multiplex real-time PCR assay. The assay was highly efficient (94.8-100.7%). It could detect fewer than 10 copies of each target sequence of the genus and each species. Twenty-five Chlamydia control and field DNA samples were differentially identified while 20 other bacterial strains comprising 10 bacterial genera were negative in the assay. This assay allows rapid, sensitive, and specific detection of the genus and the three species of avian Chlamydia in a single protocol that is suitable for routine diagnostic purposes in avian diagnostic laboratories.

Experimental inoculation of chicken broilers with C. gallinacea strain 15-56/1.

Chlamydia gallinacea is one of the new Chlamydia species, encountered predominantly in birds and occasionally in cattle, and its dissemination, pathogenicity and zoonotic potential have not yet been fully elucidated. Until now, no case of clinical infection has been described in poultry, but the number of studies is limited. This study was conducted to evaluate the course of infection and the impact on production parameters in chicken broilers inoculated with the strain 15-56/1 isolated from a Polish flock. The presence of C. gallinacea was confirmed in oropharyngeal and cloacal swabs by real-time PCR from the fifth day post inoculation (dpi). Pathogen DNA was also detected in many internal organs of inoculated chickens. All infected animals remained asymptomatic during the entire experimental period, although statistical analyses showed that broilers in the experimental group exhibited significantly lower body weight gains and feed conversion ratios than animals in the control group. These data indicate that subclinical C. gallinacea infection in broilers may lead to financial losses for poultry farmers.

Chlamydia buteonis in birds of prey presented to California wildlife rehabilitation facilities.

Chlamydial infections, caused by a group of obligate, intracellular, gram-negative bacteria, have health implications for animals and humans. Due to their highly infectious nature and zoonotic potential, staff at wildlife rehabilitation centers should be educated on the clinical manifestations, prevalence, and risk factors associated with Chlamydia spp. infections in raptors. The objectives of this study were to document the prevalence of chlamydial DNA shedding and anti-chlamydial antibodies in raptors admitted to five wildlife rehabilitation centers in California over a one-year period. Chlamydial prevalence was estimated in raptors for each center and potential risk factors associated with infection were evaluated, including location, species, season, and age class. Plasma samples and conjunctiva/choana/cloaca swabs were collected for serology and qPCR from a subset of 263 birds of prey, representing 18 species. Serologic assays identified both anti-C. buteonis IgM and anti-chlamydial IgY antibodies. Chlamydial DNA and anti-chlamydial antibodies were detected in 4.18% (11/263) and 3.14% (6/191) of patients, respectively. Chamydial DNA was identified in raptors from the families Accipitridae and Strigidae while anti-C.buteonis IgM was identified in birds identified in Accipitridae, Falconidae, Strigidae, and Cathartidae. Two of the chlamydial DNA positive birds (one Swainson's hawk (Buteo swainsoni) and one red-tailed hawk (Buteo jamaicensis)) were necropsied, and tissues were collected for culture. Sequencing of the cultured elementary bodies revealed a chlamydial DNA sequence with 99.97% average nucleotide identity to the recently described Chlamydia buteonis. Spatial clusters of seropositive raptors and raptors positive for chlamydial DNA were detected in northern California. Infections were most prevalent during the winter season. Furthermore, while the proportion of raptors testing positive for chlamydial DNA was similar across age classes, seroprevalence was highest in adults. This study questions the current knowledge on C. buteonis host range and highlights the importance of further studies to evaluate the diversity and epidemiology of Chlamydia spp. infecting raptor populations.