Genome sequence of an Acinetobacter pittii strain obtained from a red-lored parrot with pneumonia.

Acinetobacter pittii 978-A_19 was obtained from a parrot with pneumonia. It is resistant to ampicillin, carbenicillin, cephalosporins, clindamycin, and trimethoprim + sulfamethoxazole. The genome encodes a new blaADC allele, a blaOXA-502 gene, possesses several virulence genes related to adherence and biofilm formation, and has types I, II, and IV secretion systems.

Genetic diversity of Chlamydia pecorum detected in sheep flocks from Mexico.

Chlamydia pecorum, an obligate intracellular bacterium, is associated with reproductive and systemic diseases in sheep, goats, pigs, cattle, and koalas. The main conditions include polyarthritis, conjunctivitis, enteritis, pneumonia, encephalomyelitis, orchitis, placentitis, and abortion. Even though there are several studies showing that C. pecorum infections are widely spread in the world, in Mexico there are no reports. During 2016, as part of a sheep restocking program in Mexico, sheep were imported from New Zealand. Briefly after their arrival in the herds in the State of Mexico, these sheep presented abortions during the last third of gestation. A total of 62 sheep vaginal swabs that had presented abortion from different municipalities of the State of Mexico were collected. Bacterial isolation was performed using L929 mouse fibroblasts, and molecular identification was achieved by 23S rRNA (Chlamydiaceae family) and ompA gene (species-specific) real-time polymerase chain reaction (PCR). In addition, the 16S rRNA subunit and ompA gene were amplified and sequenced. Seven of 62 samples were positive for C. pecorum by bacterial isolation, 23S rRNA, and ompA gene real-time PCR. The 16S rRNA subunit and ompA gene amplicons were purified and the nucleotide sequence was determined in both directions. The consensus sequences homology search was performed using BLASTn analysis and showed a 100% of homology with the C. pecorum 16S rRNA subunit and 99% with the C. pecorum ompA gene. The population structure analyses using ompA gene demonstrated 15 genetic populations or clusters of 198 sequences from GenBank and our sequences were in a particular genetic structure corresponding to genotype "O." Herein, we describe the presence of C. pecorum in sheep imported from New Zealand into Mexico. Genetic analysis of the ompA gene showed that the isolates belong to genotype O and are related to strains isolated from sheep, cattle, and koalas.

First case of fatal equine meningoencephalitis caused by Halicephalobus gingivalis in Mexico.

Aberrant nematode larval migration in the CNS of horses is rare but frequently fatal; one of the main etiological agents involved in this illness is Halicephalobus gingivalis. This soil nematode has been associated with several fatal equine meningoencephalitis reports worldwide; however, it had never been diagnosed in horses of Mexico. A 10 year-old Andalusian horse presented dysphagia, fever, weakness, prostration and ataxia; the patient expired during the medical attention. Post mortem examination was performed and no gross alterations were found. Histopathology revealed meningoencephalitis, vasculitis and intralesional adult nematodes, larvae and eggs compatible with Halicephalobus spp. A polymerase chain reaction (PCR) for the nuclear large subunit ribosomal RNA gene (LSU rDNA) of nematodes was performed from formalin-fixed and paraffin wax-embedded sections of brain. Posterior nucleotide sequence analysis of the amplified fragment identified the agent as H. gingivalis. To our knowledge, this is the first confirmed report of Halicephalobiasis in Mexico.

Characterization of aortic and brachiocephalic filariasis by Filarioidea sp (Nematoda:Spirurida:Filarioidea) in Mexican ramphastids.

The manuscript presented herein documents the findings of filaria nematodes in 5 keel-billed toucans, and one emerald toucanet, originated from 2 private aviaries in Mexico City during two years. The birds displayed ruffled feathers, depression, inability to perch, convulsions, and sudden death. Furthermore, thickened wall of the aortic and brachiocephalic arteries, with connective tissue proliferation and chondroid metaplasia were observed. Molecular characterization matched Filarioidea sp (Nematoda: Spirurida: Filarioidea). To the authors' knowledge, this is the first documented report of filariae Filarioidea sp. causing mortality in ramphastids in Mexico. This manuscript may contribute to expand current knowledge of filariasis and the health risks and livability of wild birds.

Leptospirosis in an asplenic patient -case report.

BACKGROUND: The presentation of clinical leptospirosis has been historically associated with animal workers, slaughterhouse workers and medical veterinarians. This association has shifted to be related to flooding events and outdoor activities; few cases are related to high-risk factors found in immunosuppressed patients. Scarcely a handful of cases have serological evidence of immune response against Leptospira serovar Bratislava representing serogroup Australis, a serovar associated with poor reproductive performance in swine and horses, and recently with cats. CASE PRESENTATION: Herein, we describe a rare clinical presentation of disseminated Leptospira infection in an immunosuppressed 65-year-old woman. She was admitted to the emergency room with fever, bacteraemia, bilateral uveitis and pulmonary involvement. The patient denied outdoor activities; she only had wide exposure to faeces and urine from cats living in her home. Her medical history included idiopathic thrombocytopenic purpura (ITP) diagnosed at the age of 18. She did not respond to medical treatment, and a splenectomy was performed. At age 60, she was diagnosed with Chronic Myeloid Leukemia (CML), and was treated with a tyrosine kinase inhibitor (TKI) -Imatinib. The patient voluntarily discontinued the treatment for the last 6 months. After extensive workup, no microorganisms were identified by the commonly used stains in microbiology. The diagnosis was performed through dark-field microscopy, microagglutination test (MAT), Leptospira genus-specific PCR, the IS1500 PCR for identification of pathogenic species, and 16S based sequencing for the genus identification. CONCLUSION: Immunosuppressed patients may acquire uncommon infections from ubiquitous microorganisms. In this case, serology evidence of exposure to Leptospira serovar Bratislava by MAT and the presence of the Leptospira genus were identified. It should be on mind for the diagnosis in otherwise healthy patients, and thoroughly search on splenectomised patients exposed to animals. Additionally, this report highlights the usefulness of PCR for diagnosis of this potentially life-threatening illness.

Feline Phaeohyphomycotic Cerebellitis Caused by Cladosporium cladosporioides-complex: Case Report and Review of Literature.

Cerebellar phaeohyphomycosis was diagnosed in an 8-year-old neutered male domestic cat. Gross lesions were limited to the cerebellum, which had a focally extensive dark brown-black, soft, irregular area affecting the cortex and white matter of the left hemisphere and extending to the reticular formation. Microscopically, multifocal pyogranulomatous meningoencephalitis with intralesional pigmented fungal hyphae effaced the cerebellar grey and white matter. Fungal hyphae were 3-6 µm in diameter, septate and non-dichotomously branched, with parallel, thin and slightly bulbous walls. Polymerase chain reaction for the internal transcribed spacer 1-2 ribosomal RNA genes was performed on tissue samples from formalin-fixed and paraffin wax-embedded sections of cerebellum. Nucleotide sequence analysis of the amplified fragment identified the fungal agent as Cladosporium cladosporioides. This is the first confirmed report of cerebellar phaeohyphomycosis attributable to C. cladosporioides-complex in a domestic cat.

An impressive case of chromoblastomycosis due to Fonsecaea pedrosoi in a patient with a long history of fungal infection.

Chromoblastomycosis is defined as a chronic cutaneous and subcutaneous fungal infection caused by melanized or brown-pigmented fungi. A 63-year-old man farmer showed on external and internal part of the right arm, a well-delimited verrucous and hyperkeratotic plaque, with atrophic and cicatricial areas. Direct examination of skin scrapings samples showed the presence of muriform cells, a classic feature of chromoblastomycosis. Fungal isolation was performed in Sabouraud dextrose agar, and dark olivaceous colonies were isolated. Skin biopsy samples were obtained for histopathological and molecular diagnosis. DNA extracted from both, paraffin-embedded skin biopsy samples and fungal colonies, was used for molecular identification by 18S-ITS1-5.8S-ITS2-28S rRNA amplification and sequencing. Fonsecaea pedrosoi was identified from paraffin-embedded skin samples and fungal colonies. A combined therapy with terbinafine and itraconazole, plus cryotherapy was applied with an important improvement. Herein, we report an impressive case of chromoblastomycosis due to Fonsecaea pedrosoi with a successful outcome.

Treponema pallidum ssp. pallidum identification by real-time PCR targetting the polA gene in paraffin-embedded samples positive by immunohistochemistry.

Syphilis is a systemic and sexually transmitted infection caused by Treponema pallidum ssp. pallidum. This spirochete causes different clinical and subclinical stages depending on the duration of infection and immune status of the host. Several tests have been developed for diagnosis, and are classified into direct and indirect methods. The first one includes dark field microscopy, direct fluorescent antibody test in fluids or tissue, and molecular biology techniques. In the indirect method (serologic), the routine tests are used, and are divided in two categories: non-treponemal and treponemal ones. The objective of this work was to identify T. pallidum ssp. pallidum in paraffin-embedded skin biopsies positive by immunohistochemistry, using conventional polymerase chain reaction (PCR) and quantitative real time PCR (qPCR). We included a sample of 17 paraffin-embedded biopsies. DNA was extracted and processed by conventional PCR and real-time PCR with a TaqMan® probe to identify the polA gene. Using PCR, 11 tested positive (64.7%) and 6 (35.3%) were negative. With qPCR and TaqMan® probe, 100% of samples tested positive. The minimum number of spirochetes detected in each sample was 2. With this work, we can conclude that qPCR is a fast and very accurate method for diagnosis of syphilis in tissue specimens.

First Identification of Chlamydia psittaci in the Acute Illness and Death of Endemic and Endangered Psittacine Birds in Mexico.

A mortality episode of endemic and endangered psittacine birds from the genera Ara and Amazona occurred during January 2015. The birds were housed in a management unit for wildlife conservation that receives wild-caught birds from illegal trade. In total, 11 (57%) adult birds of different origins that shared these accommodations died. Only four of them were sent for diagnosis. The main lesions found at necropsy were consistent with those described previously for avian chlamydiosis; the presence of Chlamydiaceae was confirmed through immunofluorescence and amplification with further sequencing of the 16S ribosomal RNA gene by using hepatic tissue. Due to the lack of specific diagnostic tools on primary psittacine diseases, the pathogenic effects of systemic, respiratory, or enteric infections with high mortality rates remain unknown in Mexico. In this study, specific molecular identification of avian chlamydiosis was performed using a nested PCR on liver tissues, as well as choanal and cloacal swab samples, confirming the presence of Chlamydia psittaci in all of them. In addition, it was possible to obtain the ompA gene sequence from processed clinical samples, thereby allowing us to determine that the A genotype was affecting these birds. Although this genotype is the most commonly found worldwide in psittacine birds, this case report describes the first avian chlamydiosis outbreak affecting critically endangered and endemic psittacines subjected to reintegration programs in Mexico. Consequently, this study demonstrates the necessity of more exhaustive biosecurity strategies because other pathogens may be present and should be assessed, especially in highly threatened birds, before releasing them into their habitats.

Characterisation of quinolone-resistant Escherichia coli of 1997 and 2005 isolates from poultry in Mexico.

Forty-two enrofloxacin-resistant Escherichia coli strains isolated from eggs and first-week mortality associated with yolk sac infection of two vertically integrated poultry companies of Central Mexico in 1997 and 2005 were characterised. E. coli resistance to 19 antibiotics was determined, as well as the minimum inhibitory concentrations (broth dilution) for ciprofloxacin. The presence of gyrA,B, parC,E chromosomal point mutations, qnrA,B,S plasmid genes and the aminoglycoside acetyltransferase aac(6')-Ib-cr were determined by PCR and sequencing. Resistance to ampicillin (95%), piperacillin (95%), gatifloxacin (95%), levofloxacin (95%), ampicillin/sulbactam (90%), cefazolin (85%), trimethoprim/sulfamethoxazole (80%), amoxicillin/clavulanic acid (80%), aztreonam (80%), cefepime (80%), cefotaxime (80%), ceftazidime (80%), ceftriaxone (80%) and cefoxitin (75%) was high in the 2005 strains and 19 (95%) strains were resistant to 7 or more antimicrobials. The strains from 1997 expressed high rates of resistance only to the fluoroquinolones and 4 strains (18%) expressed resistance to 7 or more antimicrobials. All strains had a gyrA mutation (Ser83Leu) and a parC mutation (Ser80Ile or Ser80Arg) and 41 (97.6%) strains had a second gyrA mutation (Asp87Asn, Asp87Tyr or Asp87Gly). Only two (4.7%) strains had a parE mutation (Ser458Ala). A total of 10 strains were positive for the aac(6')-Ib wild-type gene, 6 strains for the aac(6')-Ib-cr variant and 6 strains possessed both the wild type and the variant. No gyrB mutations or qnrA,B,S genes were detected. This is the first report in Latin America of chromosomal and plasmid quinolone resistance genes in E. coli strains recovered from poultry.

Immunogenic response of Brucella canis virB10 and virB11 mutants in a murine model.

The virB locus, which encodes the type IV secretion system, is a major component of virulence in Brucella. A non-polar virB10 mutant and a virB11 deletion mutant were constructed in Brucella canis. In the mouse model, both mutants were cleared at day 21 post-infection, indicating reduced virulence in mice. After challenging with wild-type B. canis, the amounts of CFU recovered at day 15 were significantly lower in the group previously vaccinated with the virB10 mutant. Levels of IgG1, IgG2a, IgG2b, and IgM, the induction of the cytokines IL-2, IL-4, IL-10, and the production of IFN-γ were measured in lymphocyte cultures. All strains elicited similar levels of different antibody isotype profiles, and no significant differences were detected (P < 0.05). The wild-type strain induced a rapid and strong INF-γ response at 24 h, while both mutants induced mild INF-γ responses at 24 h, which remained constant over the course of sampling. Our results suggest that the virB mutants elicit a protective immunity and may be considered as candidates for studies to be conducted in dogs against canine brucellosis.

Characterisation of Mannheimia spp. strains isolated from bovine nasal exudate and factors associated to isolates, in dairy farms in the Central Valley of Mexico.

Mannheimia spp. strains obtained from bovine nasal exudates of either clinically healthy or clinically affected by respiratory tract disease animals were isolated and characterised to estimate the prevalence of isolated serotypes in dairy farms in Mexico, by means of a trans-sectional descriptive study. Strains were isolated and typified through biochemical and immunological tests. chi(2) or Fisher statistical tests were applied, as well as odds ratio calculation and logistic regression analysis to evaluate the association and effect of some variables on Mannheimia spp. isolation. The apparent prevalence rates of Mannheimia haemolytica was significantly higher in diseased bovines (OR = 1.94; p < 0.05), as well as in bovines younger than 1 year of age (OR = 23.98; p < 0.05), and in bovines not vaccinated against bovine pasteurellosis (OR = 1.52; p < 0.05). Age was the variable that remained in the logistic regression model. Serotype A1 showed the highest prevalence, even when most isolates were not-typable. Bovines younger than one year of age and those with disease were the groups with the highest frequency of M. haemolytica and Mannheimia glucosida isolates.

Efficacy of florphenicol premix in weanling pigs experimentally infected with Actinobacillus pleuropneumoniae.

The efficacy of a florfenicol premix was studied in weaning pigs experimentally inoculated with Actinobacillus pleuropneumoniae. Twenty five clinically healthy pigs were distributed into 3 groups; group A non-medicated, groups B and C orally medicated with 20 and 40 ppm of florfenicol respectively. The pigs were fed during 12 consecutive days and on day 5 all the groups were challenged with A. pleuropneumoniae serotype 1. All the animals in Group A developed clinical signs. Most of the pigs in the medicated groups maintained a good health status. Postmortem examination revealed severe pleuropneumonia in pigs from the control group and pneumonic lesions in 40% of the animals treated with 20 ppm of florfenicol. Development of pleuropneumonia was prevented in all the pigs medicated with 40 ppm of florfenicol. Actinobacillus pleuropneumoniae was recovered from the lungs of all control animals and from one pig of each of the medicated groups, however, the avidin biotin peroxidase (ABC-P) method detected the presence of the microorganism in all the animals. We demonstrated that medication with feed containing 40 ppm of florfenicol blocked efficiently the signs and lesions caused by A. pleuropneumoniae and increased the daily body weight gain.

Identification of four genes of the Brucella melitensis ATP synthase operon F0 sector: relationship with the Rhodospirillaceae family.

We have determined the nucleotide sequence of a cloned DNA fragment from the human and animal pathogen Brucella melitensis. Four genes were identified from a 4069 bp fragment, corresponding to the B. melitensis a, c, b', and b subunits of the ATP synthase F0 sector operon. A duplicated and divergent copy of the b-subunit gene was observed. This feature has been found only in photosynthetic bacteria and chloroplasts. In addition, the gene cluster was separated from the F1 sector, a characteristic described only for the Rhodospirillaceae family.