Molecular analysis of leptospires from serogroup Sejroe obtained from asymptomatic cattle in Rio de Janeiro - Brazil reveals genetic proximity to serovar Guaricura.

Bovine leptospirosis causes substantial reproductive failure in cattle, mainly due to infections with serovar (sv) Hardjo infection. Notwithstanding, other serovars from the serogroup (sg) Sejroe could also have important roles in bovine leptospirosis. The objective was to investigate genetic diversity of serogroup Sejroe isolates obtained from asymptomatic cattle in the state of Rio de Janeiro, Brazil. Urine and vaginal fluid (VF) were collected from clinically healthy cattle immediately after slaughter. Five isolates were recovered and characterized (serogrouping) as belonging to sg Sejroe. Sequencing of rrs and secY genes further identified them as Leptospira santarosai. Analysis of secY sequences indicated a high level of sequence homology to sv Guaricura strains. Based on culture and sequence data, we inferred that other members of sg Sejroe may be important in bovine leptospiral infection, particularly genotypes of L. santarosai serovar Guaricura.

Genotyping of Leptospira directly in urine samples of cattle demonstrates a diversity of species and strains in Brazil.

The aim of this study was to identify Leptospira in urine samples of cattle by direct sequencing of the secY gene. The validity of this approach was assessed using ten Leptospira strains obtained from cattle in Brazil and 77 DNA samples previously extracted from cattle urine, that were positive by PCR for the genus-specific lipL32 gene of Leptospira. Direct sequencing identified 24 (31·1%) interpretable secY sequences and these were identical to those obtained from direct DNA sequencing of the urine samples from which they were recovered. Phylogenetic analyses identified four species: L. interrogans, L. borgpetersenii, L. noguchii, and L. santarosai with the most prevalent genotypes being associated with L. borgpetersenii. While direct sequencing cannot, as yet, replace culturing of leptospires, it is a valid additional tool for epidemiological studies. An unexpected finding from this study was the genetic diversity of Leptospira infecting Brazilian cattle.

Molecular Characterization and Serology of Leptospira kirschneri (Serogroup Grippotyphosa) Isolated from Urine of a Mare Post-Abortion in Brazil.

A strain of Leptospira kirschneri (serogroup Grippotyphosa) was cultured from urine of a mare post-abortion in Brazil and characterized by serogrouping, multiple-locus variable-number tandem repeat analysis, PGFE, and sequencing of genes rrs and secY. Strains of L. kirschneri have apparently never been recovered from horses in tropical area, only in Europe and USA. Knowledge of local epidemiology is important to interpret genetic profiles of leptospires circulating in an area.

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.

Isolation of Leptospira interrogans Hardjoprajitno from vaginal fluid of a clinically healthy ewe suggests potential for venereal transmission.

A total of 15 adult ewes from one flock known to be seroreactive for leptospirosis was studied. Urine and vaginal fluid were collected from each animal to test for the presence of leptospires using bacterial culture and conventional PCR methods. One pure culture of Leptospira sp. was obtained from the vaginal fluid sample of a non-pregnant ewe. The isolate was characterized by DNA sequencing of the rrs and secY genes, variable-number of tandem-repeats (VNTR) analysis and serogrouping, and the isolate was typed as Leptospira interrogans serogroup Sejroe serovar Hardjo type Hardjoprajitno. This report indicates the presence of viable Leptospira in the vaginal fluid of a ewe, suggesting the potential for venereal transmission of leptospires in sheep.

Potential differences between Leptospira serovars, host-adapted (Bratislava) and incidental (Copenhageni), in determining reproductive disorders in embryo transfer recipient mares in Brazil.

The objective was to investigate potential differences between two Leptospira serovars, host-adapted (Bratislava) and incidental (Copenhageni), in causing reproductive disorders in mares. From August 2009 to March 2011, 608 adult recipient mares from eight studs located in the state of Rio de Janeiro, Brazil, were screened for leptospirosis. These mares were 3-8 years of age, of various breeds, and were managed in a semiextensive system (embryo transfer centres). According to the reproductive history of these mares, the studs were categorised as Group A (357 mares: high prevalence of reproductive problems), and Group B (251 mares: all pregnant). Of the 608 samples tested serologically, 273 (44.9 per cent) were reactive (titres ≥200), predominantly against Bratislava (62.3 per cent of reactive mares). In Group A, 247/357 sera (69.2 per cent) were reactive, with reactivity against Bratislava (64.8 per cent of reactive). By contrast, in Group B, only 26/251 sera (10.4 per cent) were reactive, the majority against Copenhageni (61.5 per cent of reactive). Seroreactivity against Bratislava was more likely to be associated (P<0.001) with reproductive problems than seroreactivity against Copenhageni; this association was true for early embryonic death (P<0.001), perinatal death (P<0.01), and abortion (P<0.001). Additionally, 29 urine samples were collected (from Group A mares) for Leptospira species PCR, of which 16 were positive (55.2 per cent). We concluded that leptospirosis was associated with poor reproductive performance in embryo recipients, with the majority due to host-adapted Bratislava.

Urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock.

The aim of the present study was to consider the wide usage of urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock. A total of 512 adult animals (300 cattle, 138 horses, 59 goats and 15 pigs), from herds/flocks with reproductive problems in Rio de Janeiro, Brazil was studied by serology and urinary PCR. From the 512 serum samples tested, 223 (43.5 %) were seroreactive (cattle: 45.6 %, horses: 41.3 %, goats: 34%and pigs: 60 %). PCR detected leptospiral DNA in 32.4 % (cattle: 21.6 %, horses: 36.2 %, goats: 77.4 % and pigs: 33.3 %. To our knowledge there is no another study including such a large number of samples (512) from different species, providing a comprehensive analysis of the usage of PCR for detecting leptospiral carriers in livestock. Serological and molecular results were discrepant, regardless the titre, what was an expected outcome. Nevertheless, it is impossible to establish agreement between these tests, since the two methodologies are conducted on different samples (MAT - serum; PCR - urine). Additionally, the MAT is an indirect method and PCR is a direct one. In conclusion, we have demonstrated that urinary PCR should be considered and encouraged as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock.

The role of horses in the transmission of leptospirosis in an urban tropical area.

The objective of this study was to demonstrate the presence of leptospires in equine urine, as evidence for a potential role of horses in transmission of this organism. Thoroughbred horses (aged 2-5 years, n = 276) from Rio de Janeiro, Brazil, were studied. After a severe storm, the premises of the animals remained flooded for 72 h. Blood samples for serology were collected on days 20 and 35 (day of storm = day 0). On day 20, 132 (47·8%) horses were seroreactive (titre ≥200) and, of these, 23 (31·0%) had increased antibody titres on day 35. Furthermore, 34 urine samples (for PCR and culture) were collected from seroreactive horses on day 35. Copenhageni was the most frequent serovar (88·8% of reactive titres). Although none of the urine samples were culture positive, 12 (35·2%) were PCR positive. This is apparently the first report of evidence of leptospires in urban horses. Furthermore, we suggest that these animals can play a role in the transmission of leptospirosis in urban areas.

Diagnosis and control of an outbreak of leptospirosis in goats with reproductive failure.

This study presents a Brazilian goat herd with reproductive failure over 2009-2010, in which there were abortions (22/50; 44%), embryonic resorption (6/50; 12%) and neonatal deaths (2/50; 4%). A diagnosis of leptospirosis was made, based on serology (microscopic agglutination test - MAT), bacterial culture, and polymerase chain reaction (PCR). Antibiotic therapy, specific vaccination protocols and changes in management practices were instigated. One year after the outbreak, diagnostic methods were repeated and reproductive performance re-analysed. Soon after the outbreak, 61/125 (48.8%) of the goats were seropositive for Leptospira. Pure isolates of Leptospira were not obtained, but Leptospira PCR testing was positive in 48/50 (96%) urine samples. After 1 year only 4.2% were seropositive and the occurrence of reproductive problems decreased roughly 10-fold, although five goats (10.4%) remained PCR-positive. A broad-based management approach, including serological and molecular diagnostic methods, vaccination, antibiotic treatment, and alteration of some environmental aspects, were critical to the control of this outbreak, thereby minimising subsequent reproductive failures and economic losses.

Assessment of the feasibility and impact of shared decision making in prostate cancer.

OBJECTIVES: To assess the feasibility and patient impact of using standardized video presentations concerning alternative treatments for managing localized prostate cancer. METHODS: One hundred eleven men with newly diagnosed localized prostate cancer were shown a video tape concerning the risks and benefits of four treatment options: radical surgery, external beam radiation, hormonal therapy, and watchful waiting. The impact of the video presentation was assessed using a questionnaire completed by the patient before and after viewing the video and again following a discussion with his treating physician. RESULTS: Patients demonstrated significant increases in their understanding of treatment options to manage prostate cancer after viewing the video presentation. Treating physicians confirmed the increased sophistication of their patients' knowledge of their disease and the potential outcomes associated with alternative treatments. CONCLUSIONS: Standardized video presentations of treatment alternatives for prostate cancer can be incorporated into busy office practices. Both patients and physicians benefit from the increased level of understanding that allows physician/patient discussions to focus on the critical risk/benefit tradeoffs rather than simply describing treatment alternatives.