Dog ectoparasites as sentinels for pathogenic Rickettsia and Bartonella in rural Guatemala.

Background: Fleas and ticks serve as vectors of multiple pathogens in the genera Rickettsia and Bartonellathat cause diseases in humans and other animals. Although human rickettsiosis and bartonellosis have been reported in all countries in Central America, limited research has been conducted to investigate the natural cycles of flea- and tick-borne rickettsiosis and bartonellosis, especially in Guatemala. Methods: We evaluated dog parasites as sentinels for zoonotic disease risk in rural Guatemala by sampling ticks and fleas from dogs, which were then identified and individually screened for Rickettsia and Bartonella. Results: A total of 77 households were surveyed and 80.52% of them had dogs. Overall, 133 dogs were examined for fleas and ticks, of which 68.42% had fleas and 35.34% had ticks. A total of 433 fleas and 181 ticks were collected from the infested dogs, with an additional 33 ticks collected from house walls. Three flea species were identified: Ctenocephalides felis (70%), Echidnophaga gallinacea(11.8%), and Pulex sp. (17.8%). Among the collected ticks, 97% were Rhipicephalus sanguineus with the rest being Amyblyomma cajennense, A. auricularium, and A. ovale. Rickettsia felis were detected in six C. felis, in one Pulex sp., and in two R. sanguineus, while Candidatus R. senegalensis was detected in one C. felis. Bartonella was detected only in fleas, including three Pulexsp. infected with B. vinsonii subsp. Berkhoffii, B. henselae, and Bartonella sp., respectively, and 11 C. felis infected with B. henselae. Conclusions: This study reports Candidatus R. senegalensis and B. vinsonii subsp. Berkhoffiiin Guatemala for the first time, and indicates the potential risk of human and dog exposure to Rickettsia and Bartonella species. These results show that dogs provide critical information relevant to managing human potential exposure to flea- and tick-borne pathogens in rural Guatemala.

Spatial distribution and population dynamics of free-roaming (stray and semi-domiciled) dogs in a major Brazilian city.

Introduction: Although estimate models have been proposed to determine free-roaming (both stray and semi-domiciled) dog populations, to date, no study has focused on the three major border areas of Brazil. Therefore, the present study assessed the free-roaming dog population of Foz do Iguaçu, a major far-west Brazilian city located in a three-border area (Brazil, Argentina, and Paraguay), which is considered among the top five Brazilian tourist destinations. Methods: Capture-release sampling was performed in three phases with a 6-month interval and 10-day duration of each phase, totaling 18 months, between 2018 and 2019. Results: A total of 1,273 dogs were estimated in the first [95% confidence interval (CI), 468-2,078 dogs], 904 in the second (95%CI, 452-1,355 dogs), and 1,564 in the third (95%CI, 521-2,607 dogs) capture phases in this area, suggesting a population density of 18.4 dogs/km2 (6.1-30.6 dogs/km2, 95% CI). Of all free-roaming dogs, 452/1,125 (40.2%) were stray with no confirmed ownership or household, whereas 672/1,125 (59.8%) had a known origin, among which 625/1,125 (55.6%) were semi-domiciled with ownership or a household, 36/1,125 (3.2%) were neighborhood dogs with maintainers, and 11/1,125 (1.0%) were owned by recycling material collectors and homeless individuals. The majority of the 1,125 dogs (862/1,125; 76.6%) had an ideal body condition score. The high outdoor access of owned dogs is likely caused by cultural behavior. However, because 533/1,125 (47.4%) of the free-roaming dogs presented with clinical abnormalities, irresponsible ownership may have negatively impacted dog health and welfare. Discussion: This study was the first to establish the density of free-roaming dogs, the ratio of stray and semi-domiciled dogs, and their dynamics over time in Foz do Iguaçu. The findings may serve as a warning for the high level of dog outdoor access and irresponsible guardianship, which may negatively affect animal health and welfare, leading to diseases, accidents, trauma, and animal cruelty.

Echocardiographic Documentation of Dilated Cardiomyopathy Development in Dogs Naturally Infected with Trypanosoma cruzi.

We aimed to characterize the echocardiographic alterations in dogs from an endemic region that were naturally infected with T. cruzi. Dogs (n = 130) seropositive for antibodies against T. cruzi and/or with acute parasitemia were enrolled in the study. Indicators of changes in the structure and systolic and diastolic functions of the left ventricle (LV) and blood flow patterns were evaluated by echocardiography. The frequency and extent of alterations in these indicators were associated with the severity of the disease. Briefly, 15 (11.54%) dogs were diagnosed with dilated cardiomyopathy (DCM), and 115 (88.46%) dogs were diagnosed as being without DCM. Infected dogs with DCM exhibited structural features of LV dysfunction, e.g., a significant (p < 0.05) increase in the LV internal diameter at systole and diastole (LVID-s, LVID-d) and a decline in the LV posterior wall (LVPW-d) thickness at diastole. Despite an increase in stroke volume and cardiac output indicating contraction force, DCM resulted in a decreased ejection fraction, affecting systolic function. Dogs that were diagnosed as DCM-negative but were positive for T. cruzi by PCR exhibited a high frequency of an increase in the thickness of the interventricular septum in systole (IVS-s) and the LV posterior wall in diastole (LVPW-d), a decline in the LV inner diameter (LVID-d, LVID-s), and fractional shortening (FS). The thinning of the LVPW at systole was the most defining feature observed in chronically infected dogs. In summary, this is the first study reporting the echocardiographic changes occurring in dogs naturally infected with T. cruzi and developing DCM. Our data suggest that changes in LVID are a major indicator of risk of cardiac involvement, and the observation of changes in the IVS, LVPW, and FS have predictive value in determining the risk of DCM development in infected dogs.

American mitogenome reference for the tropical brown dog tick, Rhipicephalus linnaei (Audouin, 1826).

The brown dog tick, Rhipicephalus linnaei (Audouin, 1826), is distributed across the American continent and is formerly known as the "tropical lineage". It belongs to the Rhipicephalus sanguineus (Latreille, 1806) species complex, referred to as R. sanguineus (sensu lato). Mitochondrial genome sequences are frequently used for the identification and represent reference material for field studies. In the present study, the entire mitochondrial genomes of R. linnaei (∼15 kb) collected from dogs in Mexico were sequenced and compared with available mitogenomes of R. sanguineus (s.l.). The mitochondrial genome is ∼90% identical to the reference genome of R. sanguineus (sensu stricto, former "temperate lineage") and > 99% identical to R. linnaei mitogenome derived from the neotype. Two additional mitogenomes were obtained and described as R. linnaei and R. turanicus from dogs in Saudi Arabia. The present study delivers a molecular reference for R. linnaei from America and complements R. linnaei mitogenomes from Africa, Asia and Australia. We propose to consider the complete mitogenome, as the reference for American R. linnaei, even when partial mitochondrial cox1, 12S rRNA or 16S rRNA genes are characterised.

Examination of the thickness of the different components of gallbladder wall in cases with mucocele and/or cholecystitis.

OBJECTIVE: To assess the thickness of each layer of the gallbladder wall with different diseases in dogs. SAMPLE: 72 gallbladders. METHODS: Retrospective study of dogs that underwent cholecystectomy. Histopathological specimens of the gallbladders were reviewed. Histopathological diagnosis was made as gallbladder mucocele or cholecystitis, and cholecystitis was further categorized into chronic cholecystitis, acute-on-chronic cholecystitis, acute cholecystitis, and necrotic cholecystitis. The thickness of each layer of the gallbladder wall was measured. RESULTS: 22 dogs were diagnosed with gallbladder mucocele without cholecystitis, 24 with gallbladder mucocele and cholecystitis, 20 with only cholecystitis, and 6 as normal. Histopathological subclassification of cholecystitis in 44 gallbladders led to diagnosis of chronic cholecystitis in 21 gallbladders, acute-on-chronic cholecystitis in 10 gallbladders, acute cholecystitis in 6 gallbladders, and necrotic cholecystitis in 7 gallbladders. The thickness of the entire wall of the gallbladder (P < .0001) and the thickness of the mucosa (P < .0001) and subserosa (P < .0001) were affected by the different disease processes. CLINICAL RELEVANCE: Layers of the gallbladder wall were affected by diseases present in the gallbladder. It resulted in a difference in the thickness of the wall of the gallbladder among the gallbladder diseases in this study. Histopathological changes should be taken into consideration before surgery while deciding what technique to use to perform a cholecystectomy.

Review of leptospirosis in dogs from Mexico: Epidemiology, diagnosis, prevention, and treatment.

Leptospirosis, classified by the World Health Organization as an emerging and neglected disease, is caused by the zoonotic pathogen Leptospira interrogans. This review aims to outline the Mexican epidemic of L. interrogans in dogs, including diagnosis and prevention methods. This review article searched articles from the publishers Wiley, Springer, PubMed, Redalyc, SciElo, and Elsevier. Among the 200 Mexican articles concerning Leptospira epidemiology, diagnosis, treatment, and vaccination, those that failed to meet the set inclusion criteria were excluded. The worldwide study of L. interrogans has focused on this bacterium. In Mexico, up-to-date information on canine prevalence, diagnosis, and vaccine use is scarce. Flow cytometrically detected Salmonella serovars differ from those in current vaccines, emphasizing the importance of broadening vaccine serovar coverage.

Infection and re-infection of Leptospira spp. in stray dogs and cats from Bogota, Colombia.

Background and Aim: Leptospirosis is a re-emerging zoonosis that is under-reported in tropical countries, and canines can be a potential reservoir of the disease. The objective of this study was to diagnose Leptospira spp. that is actively infected and re-infected in stray dogs and cats from Bogota, D.C., Colombia. Materials and Methods: A sample of 200 animals, including dogs and cats from the animal protection programs of Bogota, Colombia, were used in this study. Blood was collected from these animals for serum and DNA analysis. Conventional polymerase chain reaction (PCR) was performed using the 16s rRNA primer set, and higher-quality amplification products were sequenced by Sanger. For serodiagnosis, a group of PCR-positive samples was tested using the microagglutination test (MAT). Results: The overall PCR positivity of stray dogs and cats was 56%, 52.9%, and 65.3% in dogs and cats, respectively. The MAT seropositivity was 77.3%, and only dogs showed titers higher than 1:400. Canicola, Icterohaemorrhagiae, Pomona, Hardjo Prajitno, and Canicola and Hardjo prajitno were the serogroups associated with dogs and cats, respectively. Phylogenetic analysis revealed that the strains belonging to Leptospira interrogans serovars related to isolated samples of American, European, and Asian bats (Myotis myotis), dogs, and bovines of American origin. Conclusion: These results showed that stray dogs and cats were previously exposed to different serovars of Leptospira spp. and re-infected with other serovars that actively participated in the transmission cycle. These findings highlight the importance of actively diagnosing infectious animals to design effective intervention strategies.

Assessment and validation of the pet-owner relationship scale for Brazil.

Introduction: This study aimed to perform a cross-cultural adaptation of the cat-owner/dog-owner relationship scales. The method involved several stages: conceptual, item, semantic, operational, measurement, and functional equivalence. Procedures included translation, synthesis of translations, back-translation, consensus on the English versions, external evaluation by the original authors, expert committee evaluation, and pre-tests. Methods: The study surveyed 234 pet owners across Brazil using a 20-item questionnaire. Data analysis utilized confirmatory factor analysis, covariance-based modeling, and multigroup analysis. Results: The study confirmed the content and construct validity of the model, demonstrating good convergent validity. Hypotheses testing revealed significant inverse relationships between Perceived Cost and Perceived Emotional Closeness, and between Perceived Cost and Pet-Owner Interactions. A positive correlation was found between Perceived Emotional Closeness and Pet-Owner Interactions, with Perceived Emotional Closeness also mediating the relationship between Perceived Cost and Pet-Owner Interactions. No significant differences were found across different pet owner groups, indicating the scale's invariance and reliability across various demographics. Discussion: The study significantly expands understanding of the complex dynamics in pet-owner relationships and emphasizes the interplay between emotional and practical factors. It offers valuable insights for future research and practices in animal and human welfare.

Short-Term Effects of Two COX-2 Selective Non-Steroidal Anti-Inflammatory Drugs on the Release of Growth Factors and Cytokines from Canine Platelet-Rich Gel Supernatants.

(1) Background: There is a lack of knowledge about how a single dose of COX-2 selective non-steroidal anti-inflammatory drugs (NSAIDs) might affect the release of growth factors (GFs) and cytokines from canine platelet-rich gels (PRGs) and other hemocomponents. (2) Methods: A crossover study was conducted in six adult mongrel dogs. Animals were randomized to receive a single dose of either carprofen or firocoxib. PRG, temperature-induced platelet lysate (TIPL), chemically induced PL (CIPL), and plasma hemocomponents were obtained from each dog before (1 h) and after (6 h) the treatments. Platelet and leukocyte counts and determination of the concentrations of platelet-derived growth factor-BB, (PDGF-BB), transforming growth factor beta-1 (TGF-ß1), interleukin 1 beta (IL-1ß), tumor necrosis factor-alpha (TNF-α) and IL-10 concentrations were assayed by ELISA in all hemocomponents. (3) Results: Both platelet and leukocyte counts and PDGF-BB concentrations were not affected by NSAIDs and time. Total TGF-ß1 concentrations were not affected by NSAIDs; however, the release of this GF was increased in PRG supernatants (PRGS) at 6 h. IL-1ß and TNF-α concentrations were significantly (p < 0.001) lower in both firocoxib PRGS and plasma at 6 h, respectively. IL-10 concentrations were significantly (p < 0.001) lower at 6 h in all hemocomponents treated with both NSAIDs. (4) Conclusions: The clinical implications of our findings could indicate that these drugs should be withdrawn from patients to allow their clearance before the clinical use of PRP/PRG. On the other hand, the prophylactic use of NSAIDs to avoid the inflammatory reactions that some patients might have after PRP/PRG treatment should be performed only in those animals with severe reactive inflammation to the treatment.

Prevalence and antibiotic resistance in bacterial isolates of dogs with ulcerative keratitis in São Paulo State, Brazil.

OBJECTIVE: Identify microorganisms present in canine eyes affected by ulcerative keratitis and assess its resistance profile to available antimicrobial drugs. METHODS: Samples were collected from 88 canine eyes that exhibited ulcerative keratitis. They were identified using MALDI-TOF and subjected to antimicrobial susceptibility testing by disk diffusion. RESULTS: Among the assessed subjects, brachycephalic dogs accounted for 74.48% (50/83) of the evaluated canines. Among the 88 evaluated eyes, 90.9% (80/88) showed positive cultures, with 11.33% (10/88) of the samples isolating more than one species of bacteria. Of all bacterial isolates identified (90), Gram-positive bacteria accounted for 63.33% (57/90), while Gram-negative bacteria constituted 36.66% (33/90), with predominance of Staphylococcus spp. at 35.55% (32/90) being, Staphylococcus pseudintermedius at 68.75% (22/32), and Pseudomonas aeruginosa at 15.55% (14/90), respectively. Staphylococcus spp. exhibited resistance to penicillin (89.29%), sulfadiazine and trimethoprim (60.71%), and tetracycline (67.86%), while doxycycline (88.89%), cefotaxime (85.71%), chloramphenicol (82.14%), gentamicin, and moxifloxacin (78.57%) showed the highest sensitivity rates. Pseudomonas aeruginosa displayed sensitivity (100%) to gentamicin and imipenem, and resistance (8.33%) to norfloxacin, ciprofloxacin, and cefepime. Similarly, the Enterobacteriaceae family showed higher sensitivity to amikacin and gentamicin (88.89%), imipenem (88.24%), and levofloxacin (87.5%), with pronounced resistance to amoxicillin-clavulanate (50%) and cefazolin (47.06%). This highlights multiresistance in 23.33% (21/90) of the isolates. CONCLUSIONS: The most isolated species in canine ulcerative keratitis are S. pseudintermedius and P. aeruginosa. However, other species were also isolated, demonstrating diversity in ocular microbiota infection. There is a high-rate multidrug resistance associated with canine ulcerative keratitis. Nevertheless, these strains exhibited sensitivity to antimicrobials commonly used in veterinary ophthalmology.

Regulation of seed dormancy by histone post-translational modifications in the model plant Arabidopsis thaliana.

Plants synchronize their growth and development with environmental changes, which is critical for their survival. Among their life cycle transitions, seed germination is key for ensuring the survival and optimal growth of the next generation. However, even under favorable conditions, oftentimes germination can be blocked by seed dormancy, a regulatory multilayered checkpoint integrating internal and external signals. Intricate genetic and epigenetic mechanisms underlie seed dormancy establishment, maintenance, and release. In this review, we focus on recent advances that shed light on the complex mechanisms associated with physiological dormancy, prevalent in seed plants, with Arabidopsis thaliana serving as a model. Here, we summarize the role of multiple epigenetic regulators, but with a focus on histone modifications like acetylation and methylation, that finely tune dormancy responses and influence dormancy-associated gene expression. Understanding these mechanisms can lead to a better understanding of seed biology in general, as well as result in the identification of possible targets for breeding climate-resilient plants.

Finding Priority Areas in the Evaluation of Strategies for the Prevention of Leishmaniasis in an Endemic Municipality of Brazil.

Visceral leishmaniasis is a zoonotic disease that affects humans and dogs. The infection is endemic in the municipality of Araçatuba, São Paulo, Brazil. Given the role of dogs in the epidemiology of leishmaniasis, strategies to enhance surveillance and reduce transmission are focused on dogs. In this study, we retrospectively analyzed records of canine visceral leishmaniasis from 2013 to 2022. According to this database, the prevalence of dogs testing positive for leishmaniasis fluctuated, with an average of 65.04% (6590/10,133). Cases were clustered in 10 statistically significant areas. Environmental analyses identified a significant geographical association between animals testing positive and higher vegetation density rates compared with animals testing negative. The period from sample collection to diagnosis and euthanasia, as recommended by the Brazilian Ministry, correlated with disease prevalence and decreased over time. These findings serve to implement different action plans against leishmaniasis for each geographic region and to understand the impact and efforts of strategies in an endemic area.

Canine pulmonary clearance during feto-neonatal transition according to the type of delivery.

The success of immediate adaptation to extrauterine life depends on appropriate lung function, however, elective cesarean section can increase the risk of respiratory distress as a result of reduced pulmonary fluid absorption. This study aimed to evaluate the influence of birth mode on pulmonary clearance and respiratory performance of canine neonates in the transition period. For this purpose, 37 neonates were selected according to the obstetric condition: Vaginal Eutocia (n = 17) and Elective C-section (n = 20). Neonates were evaluated for neonatal vitality score, as well as evaluation of heart and respiratory rates, body temperature and body weight, venous hemogasometric evaluation, blood lactate and glucose, pulse oximetry and radiographic evaluation during the first 24 h of life. Additionally, amniotic fluid electrolyte composition of each puppy was evaluated. There was no influence of the type of delivery on electrolyte composition of canine amniotic fluid and neonatal pulmonary liquid content, analyzed by thoracic X-Rays. On the other hand, elective cesarean section delayed pulmonary adaptation, resulting in hypoxemia and less efficient compensatory response to acid-base imbalance and thermoregulation. In conclusion, elective c-section does not delay pulmonary clearance, whilst alters pulmonary adaptation by less efficient gas exchange and lower oxygenation, hindering the compensatory response to acid-base imbalance during the fetal-neonatal transition in dogs.

Treatment of mammary gland tumors in bitches: effects of sodium dichloroacetate as neoadjuvant therapy.

Mastectomy is the standard treatment for mammary gland tumors in dogs. In addition to traditional therapy, sodium dichloroacetate (DCA) can act as target therapy, as it may promote autophagy, reduce metastatic potential, and tumor proliferation in mammary tumor cell lines. This study aimed to analyze the effects of DCA as preoperative therapy for mammary tumors in bitches. Nineteen animals were selected, and they received DCA at a dose of 10 mg/kg orally every 12 hr, for 15 days. The periodic evaluation included hematological analysis (complete blood count and biochemical markers), evaluation of gastrointestinal adverse effects, evaluation of tumor volume, histopathological analysis, and immunohistochemical evaluation (Ki67 and cyclooxygenase-2/COX-2 markers). After treatment, there was a significant reduction in hematocrit (P=0.02) and leukocyte (P=0.04) means. Despite the variations for these two hematological parameters, the means remained within the reference range for the species. There were two cases of vomiting and one case of diarrhea. Most cases were classified as carcinoma in mixed tumor (n=7, 36.8%), followed by solid carcinoma (n=6, 31.6%). Nine cases (47.4%) showed reduced tumor volume, nine (47.4%) had stable disease, and one showed progressive disease. While there was no sample with a COX-2 score higher than 6, tumor samples with COX-2 scores 3 and 4 were significantly associated with stable disease or progression. DCA preoperative treatment for bitches with mammary gland tumors showed safety and potential cytoreduction in some cases.

Neonatal hypoglycemia in dogs-pathophysiology, risk factors, diagnosis and treatment.

Hypoglycemia is the most common metabolic alteration in the clinical routine of newborn dogs, acting as a predictor of mortality in these patients. The neonatal dog shows hepatic insufficiency and homeostatic mechanisms not yet fully developed, with limited glycogen reserves and limited capacity to perform glycogenolysis and gluconeogenesis. These physiological particularities make newborn dogs particularly susceptible to hypoglycemia when of fasting, even for short periods. Several maternal and neonatal factors may be related to a higher risk of developing hypoglycemia in neonates. This paper reviews glycemic homeostasis, the pathophysiology of neonatal hypoglycemia, the main causes involved and the diagnostic and therapeutic approaches to this condition.

Epidemiological analysis of Dirofilaria immitis (Spirurida: Onchocercidae) infecting pet dogs (Canis lupus familiaris, Linnaeus, 1758) in Baixada Fluminense, Rio de Janeiro.

Dirofilaria immitis infection is routinely detected in dogs during veterinary care in tropical and subtropical regions worldwide. Parasitological tests for the detection of this infection are routinely performed only in areas with a high prevalence. Baixada Fluminense, a region in Rio de Janeiro, was considered heartworm-free until local veterinarians began to receive blood exams results indicating the presence of microfilariae (MF). A laboratory database was hence used to collect data from 2017 to 2020 to understand the extent of spread of the parasite in this area. The results of complete blood count analysis and MF or heartworm antigen detection tests conducted on canine samples sent from veterinary clinics in Baixada Fluminense (Magé, Duque de Caxias, Guapimirim, Nova Iguaçu, and São João de Meriti municipalities) were included. In total, the results of 16,314 hematological tests were considered. The overall prevalence of D. immitis was 3.4% (554/16,314), considering that only one test result was obtained per animal on the same day. This study is highly relevant because it indicates the spreading geographic distribution of the worms, heightens awareness among local health professionals and the general population, and encourages compliance with prophylactic measures to prevent further spread of parasite.

Histopathological Analysis of Brains from Dogs Infected with Canine Distemper Virus.

We describe the use of conventional histology and immunohistochemistry against canine distemper virus (CDV) to examine the brains of domestic dogs with a confirmed diagnosis of CDV infection. Histologically, to identify the main typical lesions, we used conventional H&E stain; to evaluate the progressive demyelination, we used Luxol Fast Blue stain; and to identify the presence of viral particles in these affected regions, we used immunohistochemistry against CDV. We confirm that the histopathological analysis of brains of distemper-infected dogs is a powerful tool to evaluate the typical brain lesions and could be used as an interesting natural model to continue studying the pathogenesis of canine distemper in different species and/or other morbillivirus infections, like measles.

Life expectancy for dogs from the City of Buenos Aires.

The aim of the present study was to develop the first life tables for the dog population of the Autonomous City of Buenos Aires by constructing life expectancy tables. Data on canines received for final disposal at the Luis Pasteur Zoonosis Institute of the Autonomous City of Buenos Aires from January 2018 to December 2021 were used to prepare the life tables. Of the 11,429 dogs that died in that period, the overall life expectancy at birth was 11.88 years (95% CI = 11.37-12.39). There was no difference in life expectancy at birth by sex or by pure versus cross breeds. According to neuter status, life expectancy at birth in neutered (13.98 years) was significantly higher than in entire (11.46 years) (p-value = 0.00001). Life tables varied according to the breed studied, with the Pekingese having the highest life expectancy at birth 16.42 years (95% CI: 15.87-16.98), and the Pit bull having the lowest life expectancy at birth 10.13 years (95% CI: 9.58-10.68). The current study provides useful information for veterinary professionals and pet owners and is a valuable tool for planning and developing effective health policies.

Molecular detection and phylogenetic analysis of Borrelia Spp. In blood samples of cats and dogs by the nested-PCR method in West Azerbaijan Province, Iran.

The objective of this study was to investigate the presence and genetic attributes of Borrelia spp. in cats and dogs from the West Azerbaijan Province, located in the northwest of Iran. A total of 250 blood samples from cats and 300 blood samples from dogs were collected, and information regarding their age, sex, breed, ownership status, sampling time and region was recorded. The identification of positive samples was accomplished through nested-PCR and sequencing, with subsequent analysis of the gene sequences conducted using BioEdit software. The gene sequences for Borrelia spp. in this study showed 100% similarity to reference sequences in the GenBank® database. Phylogenetic trees were built using MEGA11. The outcomes indicated that among 250 blood samples from cats, 48 (19.2%) tested positive for Borrelia spp. gene, with a CI from 14.8 to 24.53% for cats. Similarly, out of 300 blood samples from dogs, 45 (15%) tested positive for the Borrelia spp. gene, with a CI from 11.4 to 19.48% for dogs.

The Presence of a Virulent Clone of Leptospira interrogans Serovar Canicola in Confirmed Cases of Asymptomatic Dog Carriers in Mexico.

Leptospirosis is a neglected zoonotic disease that commonly affects cattle, pigs, horses, and dogs in many countries. Infection in dogs is usually subclinical, but acute cases of leptospirosis may occur along with systemic failure, which may become fatal. After recovery from an acute infection, dogs may become asymptomatic carriers and shed pathogenic leptospires through urine for long periods of time. Here, a study of ten different cases of leptospirosis is presented, showing the relevance of dogs as asymptomatic carriers of pathogenic Leptospira. The diagnosis was confirmed via isolation and further serological and genetic identification. Four Leptospira isolates (LOCaS28, 31, 34, and 46) were obtained from the kidneys and urine samples of 58 dogs destined for destruction (6.89%) at a Canine Control Center in Mexico City. No spirochetes were observed in the urine samples of those Leptospira-positive dogs examined under dark-field microscopy, and no clinical signs of disease were observed either. Six additional isolates were obtained: two came from asymptomatic carrier dogs (CEL60 and UADY22); another isolate came from an asymptomatic dog that was a pack companion of a clinically ill dog with fatal leptospirosis (AGFA24); and finally, three isolates were taken from dogs that died of leptospirosis (LOCaS59, Citlalli, and Nayar1). Nine out of the ten isolates were identified as being from the serogroup Canicola via cross-absorption MAT using reference strains and specific antisera, and their identity was genetically confirmed as Canicola ST34 via multi-locus sequencing typing (MLST). In contrast, the isolate Nayar1 was identified as serovar Copenhageni ST2. Interestingly, the asymptomatic dogs from which Leptospira isolates were recovered consistently showed high antibody titers in the microscopic agglutination test (MAT), revealing values of at least 1:3200 against serogroup Canicola and lower titer values against other serogroups. Isolates showed different virulence levels in the hamster model. Taken as a whole, all these findings confirmed that dogs may act as asymptomatic carriers of pathogenic leptospires and possibly spread them out to the environment, thus representing an active public health risk. The results also showed that the Canicola ST34 clone is the most prevalent Leptospira serovar in dogs in Mexico, and finally that the old-fashioned MAT is a good alternative for the detection of presumptive Leptospira asymptomatic carrier dogs.