Assessment of the effect of long-term serum storage for retrospective serologic diagnosis of canine monocytic ehrlichiosis (Ehrlichia canis).

There is currently sparse information on the possible effect of long-term storage of serum specimens for the retrospective serodiagnosis of canine monocytic ehrlichiosis (CME). The aim of this study was to assess the agreement between the original serologic outcome and the results of a repeat indirect fluorescent antibody (IFA) assay for the detection of IgG antibodies against E. canis. A secondary aim was to compare the diagnostic performance of two commercially available point-of-care (POC) immunochromatographic (IC) assays. Archived serum samples originally tested as positive (n=66) or negative (n=19) for E. canis IgG antibodies and kept frozen at -20°C for a median of 22 years, were retrospectively examined by IFA and by two POC IC assays. Cohen's Kappa coefficient (0.748, p < 0.0001), indicated a substantial agreement between the original and repeat serologic testing results. An almost identical high sensitivity and moderate specificity were established for the two POC IC assays. Canine serum specimens on long-term storage may still be of value for seroepidemiologic surveys investigating the exposure to E. canis.

Orally administered gabapentin and alprazolam induce comparable levels of anxiolysis and sedation in cats.

OBJECTIVE: To assess the level of anxiolysis achieved by alprazolam and gabapentin in hospitalized cats prior to elective ovariohysterectomy and to evaluate the sedative effects of these agents. ANIMALS: 60 client-owned female cats classified as American Society of Anesthesiologists physical status 1, admitted for elective ovariohysterectomy at a veterinary teaching hospital. METHODS: The cats were prospectively and randomly allocated into 3 groups. Ninety minutes before evaluation, group G received gabapentin (100 mg/cat), group A received alprazolam (0.125 mg/cat), and group P received no medication (placebo). Stress, enclosure activity, and sedation scores were blindly evaluated. RESULTS: Stress scores were similar in cats treated with gabapentin and alprazolam and gabapentin-treated cats had significantly lower stress score than those of the placebo group. Enclosure activity levels did not differ among the groups. Additionally, gabapentin and alprazolam resulted in similar sedation levels 90 minutes after treatment, which differed significantly compared to placebo. CLINICAL RELEVANCE: The results of this study suggest that gabapentin provides similar anxiolysis in cats to that of alprazolam when evaluated 90 minutes after administration. Although no difference was noted in sedation levels between gabapentin and alprazolam, both induced deeper sedation than placebo.

Discordance between ABC blood phenotype and genotype in a domestic short-haired cat.

An adult domestic short-haired feline leukemia virus-infected cat was referred for kidney failure and worsening anemia requiring transfusions. ABC blood typing was performed with an immunochromatographic strip assay at different occasions. Gel column systems were used for the major and minor crossmatching tests, and anti-A and anti-B titers were determined. No discrete A or B bands appeared on the immunochromatographic strips at any time point for the recipient cat. The recipient's plasma agglutinated RBCs from tested type A and B cats. The recipient's RBCs appeared compatible with plasma from 1 type A and 2 B donors, and incompatible with plasma from another type A cat. Genotyping of recipient blood revealed a single homozygous c.179G>T CMAH variant predicting a blood type B. These studies suggest an unusual weak type B or missing all ABC antigens. The latter resembles the exceedingly rare Bombay phenotype in the human ABO blood group system.

Factors affecting the differential epithelial cell count in preputial cytology of healthy intact dogs.

Information is limited on the factors that may affect the preputial differential epithelial cell count in healthy intact dogs. Our objectives were to establish RIs of the preputial differential epithelial cell count in dogs and investigate the potential impact of signalment, presence of gross preputial discharge or neutrophils, preputial cleaning, and normal estrogen variation on the differential cell count. We enrolled prospectively 50 client-owned, healthy dogs in our study. Preputial cytology smears were obtained, and we performed a 4-part (parabasal, intermediate, superficial, cornified) differential cell count, before and after gentle preputial cleaning. We found higher counts of fully cornified cells (median: 11-12%) compared to previous studies. The cleaning procedure significantly decreased the intermediate (p = 0.002) and increased the superficial (p = 0.006) cells. Age was positively associated with intermediate (p = 0.007) and negatively with superficial (p = 0.01) and cornified (p = 0.02) cells. The concentration of estrogens was negatively associated with the percentage of parabasal cells (p = 0.04). In healthy dogs, a variable impact may be expected in the preputial differential epithelial cell count, by the cleaning of the prepuce before sampling, the age of the dogs, and the normal variation of estrogen concentration.

Seroprevalence of and Risk Factors for Toxoplasma gondii Infection in Cats from Greece.

Toxoplasmosis is one of the most important protozoan diseases with a global impact on the health of domestic cats and with zoonotic significance. The aims of this study were to determine the prevalence of seropositivity for Toxoplasma gondii in different populations of cats in Greece and to assess risk factors for seropositivity. A total of 457 cats were prospectively enrolled, and a commercially available indirect immunofluorescence antibody testing (IFAT) kit was used for the detection of anti-T. gondii immunoglobulin G (IgG) in serum. Overall, 95 (20.8%) of the 457 cats were seropositive for T. gondii. Based on multivariate analysis, factors associated with seropositivity included older age [Odds ratio (OR), 1.33; p < 0.001]; a history of cat-fight trauma (OR, 3.88; p = 0.004); and lack of vaccination against calicivirus, herpesvirus-1, panleukopenia, and rabies (OR, 10; p = 0.002). This study shows a high prevalence of seropositivity for T. gondii in cats in Greece. This implies that toxoplasmosis is still a major public health concern and that optimal strategies for the prevention of infection with T. gondii in cats should be established.

Investigation of comorbidities in dogs with leishmaniosis due to Leishmania infantum.

In endemic areas, dogs with leishmaniosis due to Leishmania infantum frequently have comorbidities, including mostly neoplastic, infectious, and parasitic diseases. The aim of this study was to compare the prevalence of comorbidities among dogs that are not infected by L. infantum, dogs that are infected but do not present leishmaniosis, and dogs with leishmaniosis, and to examine if certain comorbidities are independent risk factors for the infection by L. infantum and/or for the development of canine leishmaniosis (CanL). A total of 111 dogs, older than 1-year and non-vaccinated against CanL, were allocated into three groups: group A (n = 18) included dogs that were not infected by L. infantum, group B (n = 52) included dogs that were infected by L. infantum but did not present CanL, and group C (n = 41) included dogs with CanL. Signalment and historical data were obtained using a structured questionnaire. Laboratory examinations included complete blood count, serum biochemistry, urinalysis, fecal parasitology, modified Knott's test, microscopic examination of capillary blood, buffy coat, lymph node, bone marrow and conjunctival smears, qualitative serology for Dirofilaria immitis, Anaplasma phagocytophilum/A. platys, Borrelia burgdorferi and E. canis, IFAT for L. infantum, ELISA for Babesia spp. and Neospora caninum, and real-time PCR for L. infantum in bone marrow, skin biopsies and conjunctival swabs. A variety of comorbidities were found in all three groups. No independent risk factors for infection by L. infantum were found. On the contrary, among dogs infected by L. infantum, being a mongrel [odds ratio (OR): 11.2], not receiving prevention for dirofilariosis (OR: 26.5) and being seropositive to N. caninum (OR: 17.1) or to Babesia spp. (OR: 37.6), were independent risk factors for presenting CanL. Although no comorbidities influence the probability of canine infection by L. infantum, certain comorbidities may be precipitating factors for the transition from the subclinical infection by L. infantum to the overt CanL.

Seroprevalence of and risk factors for feline coronavirus infection in cats from Greece.

Feline coronavirus (FCoV) is a highly contagious and ubiquitous virus of domestic cats and wild felids. Feline infectious peritonitis (FIP) is a fatal, systemic disease caused by FCoV infection when spontaneous mutations of the viral genome take place. The aims of this study were primarily to determine the prevalence of seropositivity for FCoV in different populations of cats in Greece and assess risk factors for seropositivity. A total of 453 cats were prospectively enrolled in the study. A commercially available IFAT kit was used for the detection of FCoV IgG antibodies in serum. Overall, 55 (12.1 %) of the 453 cats were seropositive for FCoV. Based on multivariable analysis, factors associated with FCoV-seropositivity included cats adopted as strays and contact with other cats. This is the first extensive study on the epidemiology of FCoV in cats from Greece and one of the largest worldwide. Feline coronavirus infection is relatively common in Greece. Therefore, it is necessary to establish optimal strategies for the prevention of FCoV infection, considering the high-risk groups of cats identified in this study.

Clinical and Clinicopathologic Discriminators Between Canine Acute Monocytic Ehrlichiosis and Primary Immune Thrombocytopenia.

Acute canine monocytic ehrlichiosis due to Ehrlichia canis (aCME), and primary immune thrombocytopenia (pITP) are major differentials for dogs presented with thrombocytopenia, and the two diseases may clinically overlap. The aim of this study was to compare dogs diagnosed with naturally occurring aCME and pITP, to establish potentially useful clinical and clinicopathologic discriminators. A clinical record-based retrospective study was performed in 35 dogs diagnosed with aCME and 29 dogs with pITP. Dogs with aCME were significantly younger, and were more likely to experience depression or lethargy, anorexia, body weight loss, fever, lymphadenomegaly, tick infestation, and ocular discharge on admission, compared to dogs with pITP. In contrast, dogs with pITP presented more frequently with overt bleeding and had a significantly higher bleeding score compared to dogs with aCME. Dogs with aCME were more likely to be anemic and hypoalbuminemic on presentation compared to dogs with pITP. Dogs with pITP had higher white blood cell and neutrophil counts as well as lower platelet counts than dogs with aCME and were more likely to present with leukocytosis, neutrophilia and monocytosis. These clinical, hematological, and biochemical findings may be helpful discriminators between aCME and pITP, on the understanding that they will be interpreted in the context of disease-specific testing.

Prevalence and Risk Factors for Bartonella spp. and Haemoplasma Infections in Cats from Greece.

Bartonellosis and haemoplasmosis are vector-borne diseases with global impact on the health of domestic cats and of zoonotic importance. The aim of this study was to describe the epidemiological aspects of various populations of cats infected with Bartonella spp. or haemoplasma species. The populations evaluated included client-owned cats, stray cats and cats that live in breeding catteries in Greece. A total of 452 cats were prospectively enrolled into the study. A commercially available indirect immunofluorescence antibody testkit was used for the detection of Bartonella henselae IgG antibodies in serum. PCRs for the detection of Bartonella spp. and haemoplasma species DNA in the blood were also performed in a subgroup of 242 of the 452 cats. Risk factors for B. henselae seropositivity and infection with the haemoplasma species were determined using multivariable analysis. Overall, 160 (35.4%) of the 452 cats were seropositive for B. henselae. Seven (2.9%) and 46 (19%) of the 242 cats were PCR-positive for Bartonella spp. and haemoplasma species, respectively. The factors associated with B. henselae seropositivity, based on multivariate analysis, included older age, outdoor access, living region and flea infestation. Non-administration of ectoparasiticides was associated with haemoplasma species infection. This study shows a high prevalence of seropositivity for B. henselae and a relatively high prevalence of infection with haemoplasma species. Therefore, it is necessary to establish optimal strategies for the prevention of Bartonella spp. and haemoplasma species infections, considering the high-risk groups of cats identified in this study.

Prevalence of antinuclear antibodies and rheumatoid factor titers in dogs with arthritis secondary to leishmaniosis (Leishmania infantum).

Dogs with infectious arthritis may occasionally exhibit positive serum antinuclear antibody (ANA) and rheumatoid factor (RF) titers; however, relevant data are sparse for arthritis secondary to canine leishmaniosis (CanL) caused by Leishmania infantum. We determined the prevalence of positive serum ANA and RF titers in dogs with arthritis secondary to CanL. Blood samples from adult, client-owned dogs with purulent arthritis secondary to CanL, without any comorbidities, were collected for diagnostic purposes. Serum ANA titers were measured by immunoperoxidase test and RF titers by the Rose-Waaler latex test. Twelve of 23 dogs enrolled prospectively in our study had clinical arthritis, and 11 of 23 had subclinical arthritis. Based on LeishVet clinical staging, 7 dogs had clinical stage II disease, 11 had clinical stage III disease, and 5 had stage IV. None of the 23 dogs was seropositive for ANA; 3 of 23 were positive for RF. ANA and/or RF seropositivity, in dogs with CanL-associated arthritis, appears to be weak, if present at all. Based on our results, positive serum ANA and RF titers should not be expected in dogs with arthritis secondary to CanL.

Limited cross-species transmission and absence of mutations associated with SARS-CoV-2 adaptation in cats: A case study of infection in a small household setting.

In the present study, the course of SARS-CoV-2 natural infection in two asymptomatic cats, which were negative for immunosuppressive retroviral infections, is investigated. The source of the virus for the cats was their COVID-19-affected owner, with whom they were in continuous proximity in a small household setting. The owner's signs included fatigue, sneezing, anosmia and loss of taste, and diagnosis was confirmed 4 days after symptom onset. Oropharyngeal and faecal swabs were collected from the cats, to investigate the course of SARS-CoV-2 RNA concentrations, as well as the directionality of the chain of virus transmission. Both infected cats were real-time RT-PCR-positive on various time-points. Pharyngeal shedding of at least 6 days was observed in them, with high SARS-CoV-2 titres (> 7 Log10 copies/swab) on the first sampling time-point, that is, 7 days after the onset of owner's clinical signs. In one cat, after the initial decline, slightly increasing virus titres were measured 3 to 6 days after the first real-time RT-PCR-positive swab. Serological testing of this cat revealed absence of seroconversion. The course of viral RNA concentrations in the faecal swabs of the other cat was similar to that in its pharynx. The detected SARS-CoV-2 strains, from both infected cats and their owner, underwent whole-genome sequencing, revealing the absence of emergence of cross-species adaptive mutations in cats. The results support the notion that human SARS-CoV-2 strains are relatively well-adapted to cats. It is still unclear whether asymptomatic animals could play a role in COVID-19 epidemiology, in case of interaction with naïve animals and/or people. Our findings highlight difficulties in SARS-CoV-2 transmission to cats, as neither the two infected cats nor their owner was able to transmit the virus to a third cat living in the same small flat, despite their very close contact during the days corresponding to high virus shedding.

Romiplostim as Adjunctive Treatment of Refractory Amegakaryocytic Immune Thrombocytopenia in a Dog.

A 9-year-old, intact male, mixed-breed dog was admitted with a 3-day history of severe thrombocytopenia and bleeding diathesis. Physical examination revealed mucosal and cutaneous petechiae and ecchymoses, melena, and gross hematuria. Clinicopathologic evaluation indicated severe thrombocytopenia, anemia, and panhypoproteinemia. Serology for common endemic vector-borne pathogens was negative and thoracic and abdominal imaging was unremarkable. Bone marrow aspiration cytology revealed aplasia of the megakaryocytic lineage, in the context of a mildly hypoplastic myeloid and a normal erythroid series. A diagnosis of presumptive primary amegakaryocytic immune thrombocytopenia (ITP) was established. Treatment with vincristine, prednisolone, and mycophenolate mofetil along with several whole blood transfusions failed to achieve clinical and clinicopathologic remission. As an adjunct treatment, romiplostim was administered at a cumulative dose of 15 µg/kg, subcutaneously, in 2 sessions, 1 week apart, and complete clinical and hematological remission was noted 8 days postinitiation of romiplostim. Thirty-eight months later, the dog remains clinically healthy with no evidence of hematological relapse. Romiplostim could be a promising adjunctive treatment option in dogs with refractory ITP.

Presumptive Lornoxicam Intoxication in Four Dogs.

Lornoxicam is a nonsteroidal anti-inflammatory drug extensively used in human medicine, which is not approved for canine use. Lornoxicam intoxication has been rarely reported in dogs. Four dogs of various breeds, aged 7 months to 10 years, were admitted with a recent history of melena, anorexia and depression, occurring 1-4 days after the ingestion of lornoxicam (dose range: 0.53-2.7 [median 1.17] mg/kg). No clinically relevant comorbidities were documented, but low doses of prednisolone had been given in 3 of the dogs, in close temporal association with lornoxicam. Major clinical and clinicopathologic findings on admission included mucosal pallor, melena, depression, severe anemia, neutrophilic leucocytosis, and panhypoproteinemia. Perforated pyloric and duodenal ulcers were documented in 3 dogs by exploratory celiotomy or postmortem. Prolonged hospitalization (5-20 days) with extensive supportive care and multiple blood transfusions was required in 3 of the 4 dogs who survived to discharge. Lornoxicam ingestion may cause protracted and severe gastrointestinal tract injury and bleeding, blood loss anemia, panhypoproteinemia, and perforated gastrointestinal ulcers, associated with significant morbidity and mortality in dogs.

Evaluation of clinicopathological abnormalities in sick cats naturally infected by Leishmania infantum.

Feline infection by Leishmania infantum (syn. L. chagasi) has been described in areas where canine leishmaniosis is endemic. A wide variety of clinicopathological abnormalities have been reported in cats presenting clinical signs of leishmaniosis but there is a paucity of information regarding cats infected by L. infantum that do not suffer from leishmaniosis but from other diseases. The aim of this study was to compare: a) the frequency of clinicopathological abnormalities and b) the values of hematology, serum biochemistry and urinalysis parameters, between non-infected sick cats and sick cats that were infected by L. infantum. A total of 50 cats with cutaneous, ocular and/or systemic clinical signs that lived in an endemic area and had been tested for infection by L. infantum using PCR from four different tissues, were included. Based on the results of PCR, 20/50 cats were found to be infected and 30/50 non-infected. The only difference between the two groups of cats was that the concentration of inorganic phosphorus (P = 0.043) was higher in infected cats. This finding may suggest an association between infection by L. infantum and feline kidney disease.

Steroid-responsive neutropenia in a cat with progressive feline leukemia virus infection.

An 8-month-old female domestic shorthair cat was presented to the Animal Medical Center with anorexia, lethargy, and mild gastrointestinal signs. A CBC revealed a profound neutropenia, and serologic testing with an in-house test kit (SNAP FIV/FeLV Combo, IDEXX) was positive for feline leukemia virus (FeLV) antigen. Serial hematologic examinations during hospitalization showed a persistent neutropenia with occasionally severe anemia and thrombocytopenia. Prednisolone administration afforded complete hematologic remission within 3 days. Four weeks after the premature discontinuation of prednisolone, the patient relapsed; however, complete and prolonged hematologic remission was achieved after prednisolone was re-induced. Bone marrow aspiration cytology was consistent with immune-mediated destruction of the mature myeloid cells. steroid-responsive (likely immune-mediated) cytopenias rarely occur in cats with progressive FeLV infection. Although only a few cases of FeLV-positive, severely neutropenic cats that responded to immunosuppressive therapy have been reported, this case highlights that a grave prognosis should not always be given to these FeLV-positive cats.

Evaluation of serum symmetric dimethylarginine as a biomarker of kidney disease in canine leishmaniosis due to Leishmania infantum.

Canine leishmaniosis (CanL)-associated chronic kidney disease is a leading cause of morbidity and mortality in Mediterranean countries. Novel renal biomarkers, such as serum symmetric dimethylarginine (sSDMA), may be useful surrogates for the detection of renal functional impairment. The objectives of this study were to investigate sSDMA concentrations in dogs with CanL, with and without azotemia, and to establish any potential association with the prevalence and severity of proteinuria, with the prevalence of decreased urine specific gravity and with the LeishVet clinical stages of CanL. Serum samples from 68 dogs with CanL (50 nonazotemic and 18 azotemic) and 17 healthy dogs were retrospectively examined. Increased sSDMA was documented in 26 % of dogs with CanL without azotemia and in 83.3 % of dogs with azotemia. Serum SDMA was significantly higher in azotemic compared to nonazotemic dogs and was associated with the presence and severity of proteinuria, the decreased urine specific gravity and the advanced clinical stages of CanL. The results of the present study indicate that sSDMA may be a useful adjunct to serum creatinine and urine protein/creatinine ratio for the detection of CanL-associated nephropathy, but it is of limited value for distinguishing among the LeishVet clinical stages of CanL.

Capillary hemoglobin electrophoresis of healthy and anemic dogs: Quantification, validation, and reference intervals of hemoglobin fractions.

Despite the advances in canine medicine and the rapid gaining of attention of canine models in biomedical field and particularly in hemoglobin genes research, the studies on canine hemoglobin composition are sparse with ambiguous findings. Our aim was: i) to investigate the electrophoretic pattern of canine hemoglobin and the possible effects of age, sex, and anemia using a capillary electrophoresis assay, and ii) to validate this assay and calculate reference intervals (RIs) for canine hemoglobin fractions. Blood samples were collected from 53 healthy and 42 dogs with regenerative and non-regenerative anemias. The Sebia Capillarys 2 flex-piercing was used for hemoglobin analysis and it was validated using canine blood samples. R statistical language was employed for the statistical analyses. A major hemoglobin fraction (named HbA0) and a minor one (named HbA2) were identified in 100% and 47.4% of samples, respectively. The within-run and between-run CV was 0.1% for HbA0, and 9.1% and 11.2% for HbA2, respectively. The extremely narrow range of HbA0 and HbA2 values hampered a linearity study using canine blood samples. The RIs for HbA0 and HbA2 were 98.9-100% and 0-1.1%, respectively. HbA0 and HbA2 values were not significantly correlated with age (P = 0.866) or reticulocyte count (P = 0.731). No differences were observed in the median HbA0 and HbA2 between the two sexes (P = 0.887), and healthy and anemic dogs (P = 0.805). In conclusion, the capillary electrophoresis revealed a major hemoglobin fraction and an inconsistently present minor fraction. No effect of age, sex, anemia, or regenerative status of anemia was detected. The assay used was validated and RIs were generated, so as to be suitable for use in future investigations.

Congenital Unilateral Ureteral Stenosis With Hydronephrosis in a Kitten.

A 4-month-old, intact male, domestic longhair cat was admitted with a 2-month history of abdominal distention. Physical examination revealed a mild inspiratory distress and a large palpable intraabdominal mass, in an otherwise bright and alert cat. Abdominal ultrasonography revealed a large hypoechoic cystic structure compatible with hydronephrosis or a renal cyst. On exploratory celiotomy, hydronephrosis of the right kidney was documented and ureteronephrectomy was performed. Histopathology of the excised tissues revealed severe stenosis in the proximal ureteral lumen without evidence of obstructive material, a normal ureteral epithelium and severe atrophy of the renal cortex and medulla, supporting the diagnosis of congenital unilateral ureteral stenosis. The cat recovered uneventfully and 8 months later was in excellent clinical condition. Congenital ureteral stenosis with secondary hydronephrosis may occur rarely in cats and should be included in the differential diagnosis of progressive abdominal distention in young cats.

Circulating and bone marrow myeloid cells containing Leishmania amastigotes in a case of advanced canine leishmaniosis.

A 5-y-old male Poodle mix was presented with intermittent vomiting, anorexia, and weight loss. Physical examination revealed emaciation, lethargy, dehydration, hypothermia, respiratory distress, and splenomegaly. Based on clinicopathologic, serologic, and parasitologic findings, diagnoses of severe leishmaniosis and dirofilariasis were made. Extracellular, intraneutrophilic, and intramonocytic Leishmania amastigotes were observed on blood smear and buffy coat smear examination. In blood smears, 0.2% of neutrophils were observed to be infected; in buffy coat smears, 0.5% of neutrophils and 0.1% of monocytes were found to be infected. Leishmania amastigotes were also found engulfed by eosinophils and neutrophil precursors in bone marrow aspiration cytology. The detection of Leishmania amastigotes in blood smears is rare, and the clinical significance is uncertain. In circulating blood, Leishmania amastigotes are primarily found phagocytized by neutrophils. Although debatable, there is growing evidence that neutrophils are used as carriers enabling the "silent entry" of the protozoa into macrophages ("Trojan horse" theory). To date, cytologic screening of blood smears for the diagnosis of canine leishmaniosis is not a routine practice. Clinical pathologists and practitioners should be aware that Leishmania amastigotes may be present in neutrophils and less frequently monocytes during blood smear evaluation; neutrophil precursors and eosinophils may also be parasitized in bone marrow specimens.