Melissa H. Pinna

A multilocus variable number tandem repeat analysis assay provides high discrimination for genotyping Leptospira santarosai strains.

Considering the prevalence of Leptospira santarosai infections in the Americas and the scarce information about the species, we aimed to apply a multilocus variable number tandem repeat (VNTR) analysis (MLVA) for the molecular typing of L. santarosai isolates from various sources. Amplification of three VNTR loci selected from L. santarosai genome sequences resulted in a wide range of sizes for the amplified products amongst the 21 L. santarosai strains analysed. This suggested a variation in tandem repeat copy numbers in the VNTR loci. secY sequencing also showed a high nucleotide diversity, confirming the MLVA data. In conclusion, this novel MLVA provided a high level of discrimination between L. santarosai isolates, and this new typing tool could be used to investigate leptospirosis in regions where L. santarosai predominates.

Detection of Anti-Leptospira Antibodies in Captive Nonhuman Primates From Salvador, Brazil

Leptospirosis is a widely distributed zoonosis that affects several species of domestic and wild animals. Under captive conditions, Leptospirosis is a potential problem because the physical conditions in most zoos and research centers cannot prevent the captive animals from being exposed to rodents, raccoons, opossums, and other local wildlife that are known carriers. Yet, despite the potential risk, animals that are destined for reintroduction into the wild are not routinely tested for anti‐Leptospira antibodies before their release. The purpose of this study was to determine the occurrence of anti‐Leptospira antibodies in captive New World monkeys that were housed in the Wild Animals Screening Center in Salvador, Brazil. Blood samples were collected from 44 monkeys (28 Callithrix jacchus, eight Callithrix pennicilata, and eight Cebus sp.). The animals were screened for antibodies with the microscopic agglutination test. Twenty‐five (56.8%) primates were seroreactive, with Icterohaemorrhagiae being the most frequent serogroup. None of the monkeys, however, presented clinical signs of leptospirosis. Thus, seroreactivity with low titers in asymptomatic animals, as observed in this study, suggests exposure to the agent. The unexpected predominance of the serogroup Icterohaemorrhagiae further suggests that exposure to this serogroup occurred in captivity. Therefore, the dangerous possibility cannot be ignored that reintroduced monkeys will carry the leptospiral serovars into wild populations. In conclusion, primates exposed to urban serovars before their release from captivity represent a potentially significant health risk to wild populations. Am. J. Primatol. 74:8–11, 2012.

Infection by Leptospira spp. in cattle in a tropical region, Rio de Janeiro, Brazil.

Dear Sir: We read with interest the study reported by Ayral and others1 entitled “Distribution of Leptospira serogroups in cattle herds and dogs in France.” Ayral and others1 concluded that the inventory of infecting Leptospira serogroups revealed that current vaccines in France are not fully capable of preventing leptospirosis. In total, 394 cattle were diagnosed with clinical leptospirosis, and the results suggested infection by serogroups Australis (43%), Sejroe (33%), and Grippotyphosa (17%). We share our experience with bovine leptospirosis in Brazil and compare it with the scenario described in France. Infected cattle are commonly asymptomatic and may shed the bacterium in urine for long periods.2 Serology by microscopic agglutination test (MAT) is recommended as the primary diagnostic tool, although it is considered reliable only at the serogroup level.3 Although useful for a diagnosis at the herd level, serology is not adequate for the individual detection of carriers,4 which may impair control programs.5 For individuals, a rapid, sensitive, and reliable diagnostic tool, such as polymerase chain reaction (PCR), is required. In a study conducted by our group in 2013, 208 cows were randomly selected in a slaughterhouse located close to Rio de Janeiro, Brazil. After clinical pre-slaughter examination, the animals were considered free of signs of leptospirosis. Blood samples (208) for serology (MAT)3 and urine samples (199) from direct puncture of the bladder for PCR-lipL325 and culture were obtained. Serology showed that 77 of 208 (37%) animals were reactive, with serogroup Sejroe by far the most frequent (62.3%) followed by Javanica (7.8%), Icterohaemorrhagiae (6.5%), Grippotyphosa (3.9%), and Tarassovi (3.9%). Urine PCR showed that 67 of 199 animals (33.6%) were positive. Additionally, 10 isolates were obtained in pure culture, including serogroups Sejroe, Shermani, and Grippotyphosa with 2 isolates each and Sarmin, Tarassovi, Autumnalis, and Panama with 1 isolate each. These results showed a high rate of asymptomatic shedding in cows. Members of serogroups Grippotyphosa, Autumnalis, and Panama are usually associated with environmental contamination and maintained by wildlife species. Serogroup Sejroe is the most common in ruminants worldwide as well as in Brazil.6 Serogroups Sarmin, Shermani, and Tarassovi have never previously been reported from cattle in Brazil and may have been neglected as agents of bovine leptospirosis. Serogroup distribution differed between Brazil and France. In our study, both seronegative and seroreactive cows were shedding leptospires. Therefore, the direct detection of leptospires in urine by culturing or PCR is an important tool for the success of control programs in cattle. Additionally, an active surveillance approach should also include asymptomatic animals, because they may shed the bacterium to the environment and consequently, play an important role in the epidemiology of leptospirosis.

First isolation of Leptospira noguchii serogroups Panama and Autumnalis from cattle

Prevention and control of leptospirosis are based on the knowledge of locally circulating strains. Thus, efforts to obtain local isolates are paramount to the epidemiological understanding of leptospirosis. We report and discuss here the first isolation of members of serogroups Autumnalis and Panama from cattle, both belonging to Leptospira noguchii species. Urine samples (n = 167) were collected directly by puncture of the bladder from randomly selected cows from a slaughterhouse in Rio de Janeiro, Brazil, for bacteriological culture. Isolates were characterized by serogrouping and sequencing (rrs and secY genes). Overall, 10/167 positive urine samples (6%) were obtained. Sequencing of amplicons targeting for both rrs and secY genes identified two of them (2013_U73 and 2013_U232) as L. noguchii. Serogrouping of those strains indicated that 2013_U73 belonged to the Panama serogroup (titre 1600), and 2013_U232 to the Autumnalis serogroup (titre 12800). Both Panama and Autumnalis are known agents of incidental leptospirosis in cattle. This group of leptospires could be particularly important in tropical countries. This is the first report of members of serogroups Autumnalis and Panama belonging to L. noguchii species from cattle. Although related to previously reported strains, these isolates have been shown to be genetically diverse from them.

Tear production, intraocular pressure and conjunctival microbiota, cytology and histology of New Zealand rabbits (Oryctolagus cuniculus)

The purpose of this study was to establish reference values for selected ophthalmic diagnostic tests in New Zealand rabbits (Oryctolagus cuniculus). A total of 22 adult male rabbits were used. The ophthalmic tests included evaluation of tear production with Schirmer tear test 1(STT1) and Endodontic absorbent paper point tear test (EAPPTT) using two different commercial brand materials. Applanation tonometry, Culture of the conjunctival bacterial flora, , conjunctival cytology and conjunctival histology were also performed. Mean (±SD) for STT1, EAPPTTa, EAPPTTb and IOP was 7.27±2.51mm/min, 12.43±1.69mm/min, 15.24±2.07mm/min, 12.89±2.80mm Hg, respectively. Staphylococcus epidermidis, Staphylococcus sp. and Bacillus sp. were predominant. The cytological evaluation revealed the presence columnar epithelial cells, superficial squamous keratinized cells, lymphocytes, heterophils, red blood cells, mucus and bacteria. The histological analysis revealed a stratified epithelium, characterized by the presence of columnar epithelial cells with a large number of goblet cells. The reported data can be used for therapeutic or experimental purposes.

Conjunctival flora, Schirmer's tear test, intraocular pressure, and conjunctival cytology in neotropical primates from Salvador, Brazil

This study aimed to establish reference values for selected ophthalmic diagnostic tests in healthy neotropical primates from Salvador, Brazil.

Exophthalmos due to odontogenic intraorbital abscess in Cebus apella

The accumulation of pus in the orbit originating from an infected dental root is classified as odontogenic intraorbital abscess.

Detection of bovine carriers of Leptospira by serological, bacteriological, and molecular tools

Bovine leptospirosis is an important infectious disease that causes reproductive problems and economic risks, particularly in the tropics. The present study aimed to determine the extent of Leptospira infection among bovines on a slaughterhouse from Rio de Janeiro, Brazil via serological, bacteriological, and molecular tests. Two hundred eight bovines were examined in total, and we obtained 208 blood samples for serology, 198 urine samples collected via direct bladder puncture for polymerase chain reaction (PCR) and culture, 208 kidney samples (one from each animal) for PCR and culture, and 92 vaginal fluid samples from sterile swabs for PCR and culture. Serology demonstrated that 77/208 (37%) of the animals presented anti-Leptospira antibodies. Serogroup Sejroe was by far the most common. One hundrd thirty-three animals (63.9%) were PCR positive in at least one of the tested samples and were considered as Leptospira carriers. Furthermore, ten isolates were obtained by pure culture, all of them from urine samples. Bovine leptospirosis is widely prevalent, and the occurrence of renal carriers was unexpectedly much higher than generally reported.

Conjunctival bacterial flora and antimicrobial susceptibility of captive and free-living sea turtles in Brazil

PURPOSE To describe the aerobic conjunctival bacterial flora of 3 especies of free-living and under human care sea turtles and determine its antimicrobial susceptibility in vitro. METHOD Thirty-six sea turtles (72 eyes), juveniles and adults, 7 free-living Chelonia mydas and 8 Chelonia mydas, 4 Caretta caretta, 11 Eretmochelys imbricata, and 6 Lepidochelys olivacea under human care, were evaluated. Conjunctival cultures were collected for identification of aerobic bacteria and antimicrobial susceptibility testing for ciprofloxacin, chloramphenicol, gentamicin, neomycin, oxacillin, polymyxin B, tetracycline, and tobramycin using antibiotic disks. Bacterial strains showing no sensitivity to 4 or more antimicrobials were considered multiresistant to this panel. RESULTS Bacterial growth was observed in 12/14 (85.71%) samples in the free-living sea turtles, and there was growth in 100% (58/58) of the samples from captive animals. There were 94 strains isolated and 15 species identified. There was a predominance of Gram-positive bacteria in free-living Chelonia mydas, most of which were Bacillus and Staphylococcus. The most commonly isolated Gram-negative species were enterobacteria for free-living and under human care animals. The strains were predominantly sensitive to ciprofloxacin and tobramycin, and less sensitive to oxacillin or polymyxin B. Ten multiresistant strains were isolated. Yeast were identified in 13.89% (10/72) of the samples. CONCLUSIONS These results, showing differences in the conjunctival bacterial flora of free-living and captive animals, may be helpful for diagnosis and treatment of ocular disorders in sea turtles.

Indícios de exposição a leptospiras em primatas neotropicais resgatados do comércio ilegal e de um Zoológico da Bahia

Few studies have compared the seroprevalence of antileptospiral agglutinins with the demonstration of urinary shedding of leptospires or evidence of active infection in the bloodstreams of non-human primates. The study population consists of 58 animals, including d 42 monkeys from the Zoological Park of Salvador (Parque Zoobotânico Getulio Vargas), Bahia, Brazil. The study also evaluated 16 primates (Cebus sp.) rescued from illegal trade that were housed in the Wildlife Rehabilitation Center of Salvador (CETAS), Bahia, Brazil. The seroprevalence of antileptospiral antibodies was low (2%) in the animals from the Zoo. A higher rate (31%) was observed among the animals that were rescued from illegal trade in the state of Bahia. Even if all the blood and urine samples were negative for leptospiral DNA fragments, the high frequency of serological evidence of exposure suggests a potential risk of leptospirosis transmission when keeping these animals as pets.