Clinical and Laboratory Profile of Dogs Seroreactive to Ehrlichiosis Treated at the Veterinary Medical Teaching Hospital in Niterói, State of Rio de Janeiro, Brazil

Background: Ehrlichiosis is a tick-borne disease highly prevalent in Brazil, and is relevant in canine clinical practice due to its high morbidity and mortality. Its clinical signs are nonspecific and its phases are acute, lasting 2 to 4 weeks; subclinical, i.e., asymptomatic; and chronic, resembling an autoimmune disease. The purpose of this study was to identify the occurrence of reactivity to Ehrlichia canis of bitches treated at the Veterinary Medical Teaching Hospital of the Universidade Federal Fluminense (UFF) - Niterói, RJ, Brazil, based on serological examination by iELISA, and to compare the hematological, biochemical, urinary protein-creatinine and urinary density profiles of reactive and non-reactive animals. Materials, Methods & Results: This study involved solely bitches, regardless of breed, starting at 1 year of age. One hundred and thirty bitches, 1 to 16 year-old (mean age 7.02 ± 4.00), weighing 1.5 to 50 kg (mean weight 12.12 ± 10.65) were subjected to clinical examination and abdominal ultrasound. Complete blood count, biochemical measurements, urinalysis and serology for E. canis were also performed. The serum was used in the iELISA to identify immunoglobulin G (IgG), using a canine Ehrlichia Imunotest® diagnostic kit (Imunodot®, Jaboticabal, SP, Brazil) according to the manufacturer’s instructions. Sixty animals (46.20%) were reactive to E. canis. According to their owners, only 5 (8.3%) of the 60 seroreactive animals had a history of tick-borne disease. The most common profile was that of mixed breed animals living with their owners, older than 7 years, who had not been treated preventatively with specific drugs against ectoparasites. Laboratory tests showed significant differences between groups in terms of total protein (TP), and calcium and urinary protein-creatinine ratio (UPC). TP and UPC were elevated in the non-reactive group, while the only significant change in the reactive group was mild hypocalcemia. In this study, 30% (18/60) of the bitches were seroreactive to E. canis and had hypocalcemia. Of these, 50% (9/18) had a UPC above 0.5. Furthermore, 66.7% (12/18) of this group with hypocalcemia also showed urine density (UD) of less than 1024. Among these 18 bitches, 5 had both alterations, i.e., UPC > 0.5 and UD < 1024.Discussion: In this study, a high prevalence of bitches seroreactive to Ehrlichia canis was observed, despite the absence of clinical and/or laboratory signs indicative of the disease. In the investigation of IgG class antibodies, it is not possible to determine the exact time of infection, and titers may remain high for a period of more than 11 months, even after treatment and elimination of the bacterium. The fact that most seroreactive bitches showed no symptoms compatible with the disease either before or during the study suggests that they were in the subclinical phase of ehrlichiosis. The main reason for calcium metabolism disorders is a phosphorus imbalance, a condition that occurs in kidney diseases. Isosthenuria reflects the kidney’s inability to concentrate urine. This finding may be one of the first clinical manifestations of chronic kidney disease (CKD), especially in dogs. On the other hand, the UPC ratio may increase with the progression of CKD. The presence of hypocalcemia, isosthenuria and increased UPC associated with seroreactivity suggests that infection by E. canis may be associated with the onset of CKD. Veterinarians should keep in mind the complexity of the pathophysiology of ehrlichiosis to ensure the disease is not underdiagnosed in any of its phases, thereby ensuring the correct treatment is provided. Such awareness is expected to reduce the chronicity of the disease and underlying sequelae among dogs.Keywords: Ehrlichia canis, serology, tick, clinic, renal.Descritores: Ehrlichia canis, sorologia, carrapato, clínica, renal.Título: Perfil clínico e laboratorial de cadelas sororeativas para erliquiose tratadas em um Hospital Veterinário Universitário em Niterói, Estado do Rio de Janeiro, Brasil

Investigação de Borrelia spp. e alterações hematológicas associadas em Gambás (Didelphis aurita e Didelphis albiventris) de vida livre no Estado do Rio de Janeiro

A Síndrome Baggio-Yoshinari (SBY) ou Borreliose brasileira é uma zooonose causada por um grande número de espécies do complexo Borrelia burgdorferi sensu lato. No Brasil, há relatos de soropositividade em seres humanos, animais domésticos e silvestres. Marsupiais do gênero Didelphis (Linnaeus, 1758) são mamíferos com ampla distribuição nas Américas que apresentam hábito peridomiciliar, o que aumenta o risco de introdução e dispersão de agentes patogênicos. Diante disso, o objetivo do estudo foi investigar ocorrência de Borrelia spp. em Didelphis aurita e D. albiventris de vida livre, que habitavam o bioma Mata Atlântica do Rio de Janeiro, e correlacionar os achados hematológicos com a infecção. Foram analisadas 60 amostras sanguíneas de gambás das espécies D. aurita e D. albiventris, independente de sexo, idade e estado clínico, resgatados pelo CRAS – UNESA e atendidos na Universidade de Vassouras. A análise hematológica foi realizada por método manual e a análise molecular por nested-PCR, utilizando primers específicos. Das 60 amostras analisadas, 23,33% (14/60) foram positivas para Borrelia spp. Das sequências de DNA obtidas nesse estudo, 85,71% (12/14), apresentaram alta similaridade (94-99%) com as sequências de Borrelia burgodoferi (8/12 – 66,67%) e Borrelia sp. (4/12 – 33,33%) disponíveis no GenBank. Não houve diferença significativa nos parâmetros hematológicos entre animais positivos e negativos e não foi encontrado espiroquetas nos esfregaços sanguíneos avaliados. Conclui-se que marsupiais podem participar da epidemiologia da doença de Lyme no Brasil e a detecção de Borrelia spp. em D. aurita e D. albiventri confirma que a bactéria habita a Mata Atlântica do Estado do Rio de Janeiro, indicando a manutenção de um ciclo silvestre desse patógeno. Ainda, a ausência de alterações clínicas e hematológicas entre animais positivos, sugere a atuação desse marsupial como reservatório de Borrelia spp.Baggio-Yoshinari Syndrome (BYS) or Brazilian Borreliosis is a zoonosis caused by Borrelia burgdorferi sensu lato complex. In Brazil, there are reports of seropositivity in humans, domestic and wild animals. Didelphis (Linnaeus, 1758) are mammals with wide distribution in the Americas that present peridomiciliary habit, which increases the risk of introduction and dispersion of pathogenic agents. Therefore, the aim of this study was to investigate the occurrence of Borrelia spp. in the free-living Didelphis aurita and D. albiventris, who inhabited Atlantic Forest biome of Rio de Janeiro, and to correlate hematological findings with infection. Sixty blood samples from D. aurita and D. albiventris opossums were analyzed, regardless of sex, age and clinical status, rescued by CRAS – UNESA and attended at Universidade de Vassouras. Hematological analysis was performed by manual method and molecular analysis by nested-PCR, using specific primers. Of the 60 samples analyzed, 23.33% (14/60) were positive for Borrelia spp. Of the DNA sequences obtained in this study, 85.71% (12/14) showed high similarity (94-99%) with the sequences of Borrelia burgodoferi (8/12 – 66.67%) and Borrelia sp. (4/12 – 33.33%) available on GenBank. There was no significant difference in hematological parameters between positive and negative animals and no spirochetes were found in the blood smears evaluated. It is concluded that marsupials can participate in the epidemiology of Lyme disease in Brazil and the detection of Borrelia spp. in D. aurita and D. albiventris confirms that the bacterium inhabits Atlantic Forest of Rio de Janeiro State, indicating the maintenance of a wild cycle of this pathogen. Furthermore, the absence of clinical and hematological alterations among positive animals suggests the role of this marsupial as a reservoir of Borrelia spp.59f

O escarro induzido no diagnóstico das doenças pulmonares em pacientes positivos ao vírus da imunodeficiência humana

INTRODUÇÃO: O escarro induzido é utilizado para monitorar a inflamação de vias aéreas, porém seu papel como ferramenta diagnóstica de doenças pulmonares em pacientes imunocomprometidos ainda necessita de melhor definição. OBJETIVOS: Determinar o rendimento do escarro induzido no diagnóstico das doenças pulmonares em pacientes positivos ao virus da imunodeficiência humana (HIV). MÉTODO: No período de janeiro de 2001 a setembro de 2002, foram avaliados todos os pacientes com idade superior a 14 anos, infectados com o HIV, admitidos em Hospital de Referência. Foram incluídos aqueles indivíduos que apresentavam manifestações clínicas do aparelho respiratório há pelo menos 7 dias, associadas, ou não, a alterações radiológicas, bem como indivíduos assintomáticos do ponto de vista respiratório, com alterações no radiograma de tórax. Os pacientes foram submetidos à avaliação clínica, radiológica e laboratorial e realizaram a indução de escarro, seguida pela broncofibroscopia, lavado broncoalveolar e biópsia pulmonar transbrônquica. As amostras foram processadas para bacterioscopia pelo método de Gram e Ziehl-Neelsen, cultura quantitativa para bactérias, exame micológico direto, cultura para micobactérias e fungos, pesquisa de citomegalovírus e Pneumocystis jiroveci, bem como celularidade total e diferencial. RESULTADOS: 54 pacientes foram incluídos no estudo. A pesquisa de agente etiológico resultou negativa em 7 pacientes, sendo que nos casos restantes foram isolados 60 agentes. Dentre os agentes isolados, 46,7% foram P. jiroveci; 33,5% bactérias piogênicas e 16,7% Mycobacterium tuberculosis. O escarro induzido apresentou sensibilidade de 57,5%, especificidade de 42,9%, valor preditivo positivo de 87,1%, valor preditivo negativo de 13% e acurácia de 55,6%. CONCLUSÕES: Nesta população, a análise do escarro induzido é um procedimento simples, seguro e com bom rendimento diagnóstico

Exacerbations of COPD and symptoms of gastroesophageal reflux: a systematic review and meta-analysis

OBJECTIVE: To examine the relationship between gastroesophageal reflux (GER) and COPD exacerbations. METHODS: We conducted a systematic search of various electronic databases for articles published up through December of 2012. Studies considered eligible for inclusion were those dealing with COPD, COPD exacerbations, and GER; comparing at least two groups (COPD vs. controls or GER vs. controls); and describing relative risks (RRs) and prevalence ratios-or ORs and their respective 95% CIs (or presenting enough data to allow further calculations) for the association between GER and COPD-as well as exacerbation rates. Using a standardized form, we extracted data related to the study design; criteria for GER diagnosis; age, gender, and number of participants; randomization method; severity scores; methods of evaluating GER symptoms; criteria for defining exacerbations; exacerbation rates (hospitalizations, ER visits, unscheduled clinic visits, prednisone use, and antibiotic use); GER symptoms in COPD group vs. controls; mean number of COPD exacerbations (with symptoms vs. without symptoms); annual frequency of exacerbations; GER treatment; and severity of airflow obstruction. RESULTS: Overall, GER was clearly identified as a risk factor for COPD exacerbations (RR = 7.57; 95% CI: 3.84-14.94), with an increased mean number of exacerbations per year (mean difference: 0.79; 95% CI: 0.22-1.36). The prevalence of GER was significantly higher in patients with COPD than in those without (RR = 13.06; 95% CI: 3.64-46.87; p < 0.001). CONCLUSIONS: GER is a risk factor for COPD exacerbations. The role of GER in COPD management should be studied in greater detail

Pelger-Huёt Anomaly in a Bitch Basenji

Background: Pelger-Huët anomaly (PHA) is characterized by morphological changes in all granulocytes, being more evident in neutrophils. Granulocytic function in these animals remains unchanged. Hereditary form of PHA should be differentiated from pseudo-PHA,a condition acquired from infections and/or inflammation conditions. Recognition of PHA is important to avoid misleading leukogram interpretation, since hyposegmentation of neutrophils can be confused with left shift, making it necessary to carry out diagnostic tests and treatment for the disease that is generating the deviation. The objective of this case report was to demonstrate the importance of laboratory diagnosis in PHA. Case: A 11-month-old bitch Basenji, was presented to perform preoperative evaluation for elective ovariosalpingohisterectomy at Veterinary Hospital (HUVET) of the Universidade Federal Fluminense (UFF). Tutor reported that animal was healthy, vaccination status was current, had deworming protocol applied and had not made use of medications recently. Animal presented normophagy, normodipsia, normuria and normochezia. Upon physical examination, animal was alert consciousness level, with adequate hydration, hyperemic oral mucosa, a less than 2 s capillary perfusion time, normal lymph nodes (submandibular, pre-scapular, inguinal and popliteal), rectal temperature of 39.2°C, heart rate of 160 beats per minute and respiratory rate of 60 movements per minute, possibly due to the animal’s agitated state. Abdominal palpation showed no changes. Physical examination presented no alterations. Preoperative exams included complete blood count (CBC) and biochemistry profile (ALT, AP, glucose, creatinine, urea, total proteins and fractions). Samples were sent to Hospital's Veterinary Clinical Pathology Laboratory (LABHUVET/UFF) for analysis. CBC was performed using automatic method. Blood smears were stained with hematological stain and then a cytomorphological evaluation was performed. The first CBC revealed 23% of neutrophils with nuclear hyposegmentation and 44% of neutrophils were bandsA follow up was performed after 9 months, and a Complete Blood Count was performed again in which 12% of neutrophils showed nuclear hyposegmentation with mature chromatin pattern, 40% of neutrophils were bands, 1% of meta-myelotcytes neutrophils, 1% of myelocytes neutrophils and, also, eosinophils with nuclear hyposegmentation. Animal was healthy, and had no alterations on physical examination suggesting a diagnosis of PHA. Discussion: Recognition of PHA is important to avoid misleading leukogram interpretation, since neutrophils hyposegmentation can be confused with left shift, which is considered severe with poor prognosis, making it necessary to carry out diagnostic tests and treatment for the disease that is generating the deviation. The diagnosis of PHA was considered by the shape of the leukocytes nuclei, without evidence of inflammatory disease, during the patient follow up. Therefore, this anomaly should be considered as a differential diagnosis of left shift, thus avoiding unnecessary clinical and therapeutic procedures. Guidance in face of this hereditary hematological syndrome is important. The responsible guardian of the animal must not allow it to act as a breeder in order to interrupt possible transmission of this anomaly to offspring, because there is a fatal form when it comes to homozygotes. Keywords: canine, dog, hereditary anomaly, WBC, nuclear hyposegmentation, neutrophils, left shift

Pelger-Huёt Anomaly in a Bitch Basenji

Background: Pelger-Huët anomaly (PHA) is characterized by morphological changes in all granulocytes, being more evident in neutrophils. Granulocytic function in these animals remains unchanged. Hereditary form of PHA should be differentiated from pseudo-PHA,a condition acquired from infections and/or inflammation conditions. Recognition of PHA is important to avoid misleading leukogram interpretation, since hyposegmentation of neutrophils can be confused with left shift, making it necessary to carry out diagnostic tests and treatment for the disease that is generating the deviation. The objective of this case report was to demonstrate the importance of laboratory diagnosis in PHA. Case: A 11-month-old bitch Basenji, was presented to perform preoperative evaluation for elective ovariosalpingohisterectomy at Veterinary Hospital (HUVET) of the Universidade Federal Fluminense (UFF). Tutor reported that animal was healthy, vaccination status was current, had deworming protocol applied and had not made use of medications recently. Animal presented normophagy, normodipsia, normuria and normochezia. Upon physical examination, animal was alert consciousness level, with adequate hydration, hyperemic oral mucosa, a less than 2 s capillary perfusion time, normal lymph nodes (submandibular, pre-scapular, inguinal and popliteal), rectal temperature of 39.2°C, heart rate of 160 beats per minute and respiratory rate of 60 movements per minute, possibly due to the animal’s agitated state. Abdominal palpation showed no changes. Physical examination presented no alterations. Preoperative exams included complete blood count (CBC) and biochemistry profile (ALT, AP, glucose, creatinine, urea, total proteins and fractions). Samples were sent to Hospital's Veterinary Clinical Pathology Laboratory (LABHUVET/UFF) for analysis. CBC was performed using automatic method. Blood smears were stained with hematological stain and then a cytomorphological evaluation was performed. The first CBC revealed 23% of neutrophils with nuclear hyposegmentation and 44% of neutrophils were bandsA follow up was performed after 9 months, and a Complete Blood Count was performed again in which 12% of neutrophils showed nuclear hyposegmentation with mature chromatin pattern, 40% of neutrophils were bands, 1% of meta-myelotcytes neutrophils, 1% of myelocytes neutrophils and, also, eosinophils with nuclear hyposegmentation. Animal was healthy, and had no alterations on physical examination suggesting a diagnosis of PHA. Discussion: Recognition of PHA is important to avoid misleading leukogram interpretation, since neutrophils hyposegmentation can be confused with left shift, which is considered severe with poor prognosis, making it necessary to carry out diagnostic tests and treatment for the disease that is generating the deviation. The diagnosis of PHA was considered by the shape of the leukocytes nuclei, without evidence of inflammatory disease, during the patient follow up. Therefore, this anomaly should be considered as a differential diagnosis of left shift, thus avoiding unnecessary clinical and therapeutic procedures. Guidance in face of this hereditary hematological syndrome is important. The responsible guardian of the animal must not allow it to act as a breeder in order to interrupt possible transmission of this anomaly to offspring, because there is a fatal form when it comes to homozygotes. Keywords: canine, dog, hereditary anomaly, WBC, nuclear hyposegmentation, neutrophils, left shift