Infecção por Anaplasma platys em um canino da raça poodle: relato de caso / Anaplasma platys infection in a poodle canine: case report / Infección por Anaplasma platys en un canino de raza caniche: reporte de caso

Um canino da raça Poodle, 15 anos, macho, inteiro, foi atendido no Hospital Veterinário da Universidade Luterana do Brasil apresentando: hiporexia há cinco dias e anorexia nos dois últimos dias, hipodipsia, prostração, hipertermia, uveíte e um episódio de vômito. Ao exame físico, constatou-se 8% de desidratação, mucosas secas e hipocoradas, tempo de preenchimento capilar maior que dois segundos e presença de carrapatos (Rhipicephalus sanguineus). Devido à presença dos carrapatos, foi administrado afoxolaner. Realizou-se o exame parasitológico de sangue (EPS), colhido da ponta da orelha e o teste rápido (4Dx® IDEXX). Com o resultado positivo do teste rápido e visualização de mórula no interior das plaquetas compatíveis com Anaplasma platys, definiu-se o diagnóstico de anaplasmose trombocítica canina. A partir do diagnóstico, instituiu-se o tratamento com doxiciclina a cada 12 horas por via oral e, após 48 horas de tratamento, o cão recebeu alta e seguiu com tratamento em domicílio. Após 21 dias de antibioticoterapia, o paciente apresentou melhora clínica satisfatória. Realizou-se nova coleta para EPS, que apresentou resultado negativo.

A 15-year-old male Poodle canine was attended at the Veterinary Hospital of the Lutheran University of Brazil presenting: hyporexia of five days and anorexia for the last two days, hypodipsia, prostration, hyperthermia, uveitis and one episode of vomiting. Physical examination revealed 8% dehydration, dry and pale mucous membranes, capillary refill time greater than two seconds and the presence of ticks (Rhipicephalus sanguineus). Due to the presence of ticks, afoxolaner was administered. The parasitological blood test (EPS) was performed, collected from the ear vein, and a snap test (4Dx® - IDEXX). The positive result of the snap test and visualization of the morulae inside the platelets compatible with Anaplasma platys, the diagnosis of canine thrombocytic anaplasmosis was reached. The treatment with doxycycline was instituted every 12 hours orally and after 48 hours the dog was discharged and continued with treatment at home. After 21 days of antibiotic therapy, the patient showed satisfactory clinical improvement. A new collection for EPS was performed, which showed a negative result.

Un canino macho de 15 años, entero, fue atendido en el Hospital Veterinario de la Universidade Luterana do Brasil por presentar: hiporexia durante cinco días y anorexia durante los dos últimos días, hipodipsia, postración, hipertermia, uveitis y un episodio de vómitos. Al examen físico se encontró un 8% de deshidratación, mucosas secas y pálidas, tiempo de llenado capilar superior a dos segundos y presencia de garrapatas (Rhipicephalus sanguineus). Debido a la presencia de garrapatas, se administró afoxolaner. Se realizó el examen parasitológico de sangre (EPS), recogida de la punta de la oreja y la prueba rápida (4DX - IDEXX). Con el resultado positivo de la prueba rápida y visualización de mórula en el interior de las plaquetas compatible con Anaplasma platys, se definió el diagnóstico de anaplasmosis trombocítica canina. Tras el diagnóstico, se instauró tratamiento con doxiciclina cada 12 horas por vía oral y, a las 48 horas de tratamiento, se dio de alta al perro y se continuó con el tratamiento en casa. Después de 21 días de antibioticoterapia, la paciente mostró una mejoría clínica satisfactoria. Se realizó un nuevo cobro de EPS, que arrojó un resultado negativo.

Aspectos epidemiológicos e hematológicos de cães infectados com Ehrlichia sp e Anaplasma sp em Jataí - GO, Brasil / Epidemiological and hematological aspects of dogs with Ehrlichia sp and Anaplasma sp in Jataí - GO / Aspectos epidemiológicos y hematológicos de perros con Ehrlichia sp y Anaplasma sp en Jataí - GO

No Brasil, as infecções por hemoparasitas representam uma das principais causas de busca por assistência veterinária em cães. Infecções por bactérias da ordem Rickettsiales que incluem Ehrlichia sp. e Anaplasma sp.tem uma grande participação nessas buscas. Foi objetivo deste estudo elucidar aspectos epidemiológicos regionais e hematológicos de infecções por Anaplasma sp.e Ehrlichia sp, muitas vezes assintomáticas, em cães no município de Jataí-GO. Os dados deste estudo foram baseados em amostras de sangue de cães atendidos no Hospital Veterinário da Universidade Federal de Jataí, em Jataí, Goiás, entre os anos de 2005 e 2017. A observação microscópica da presença de mórulas de Ehrlichia sp. e Anaplasma sp. foi utilizada como critério de positividade. 5.015 animais foram atendidos nesse período e 691 (13,78%) deles foram positivos para a presença destes hemoparasitas. Entre esses animais, 9,23% (463/5015) possuíam mórulas de Ehrlichia sp., e 4,37% (219/5015) apresentavam mórulas típicas de Anaplasma sp.,enquanto 0,18% (9/5015) possuíam mórula de ambas as espécies. As fêmeas representaram 57,89% (400/691) dos animais positivos e os machos 42,11% (291/691). 35,02% (242/691) dos animais tinham menos de 1 ano de vida, 54,13% (374/691) tinham entre 1 e 8 anos, e 10,85% (75/691) tinham mais de 8 anos. As fêmeas, com menos de um ano de idade e coinfectadas, apresentaram as alterações mais significativas nos aspectos hematológicos, hematócrito (29%), contagem de hemácias (3 milhões/mm3), contagem de plaquetas (40.000/mm3) e contagem total de leucócitos (16.000 mil/mm3), evidenciando anemia e trombocitopenia. Animais infectados com Anaplasma sp.e Ehrlichia sp.,ao mesmo tempo, apresentaram trombocitopenia mais intensa, evidenciada em machos entre 1 e 8 anos (30.000/mm3). As infecções por Anaplasma sp. causaram uma diminuição severa na média da contagem de glóbulos vermelhos, evidenciada em machos (3 milhões/mm3) e fêmeas com menos de 1 ano (3,79 milhões/mm3).

In Brazil, infections by hemoparasites represent one of the main causes of seeking veterinary assistance in dogs. Infections with bacteria of the order Rickettsiales that include Ehrlichia sp. and Anaplasma sp. has a great participation in these assistances. The objective of this study was to elucidate regional epidemiologic and hematologic aspects of infections by Anaplasma sp. and Ehrlichia sp.,often asymptomatic, in dogs in the municipality of Jataí-GO. The data in this study were based on blood samples from dogs treated at the Veterinary Hospital of the Federal University of Jataí, in Jataí, Goiás, between the years 2005 and 2017. Microscopic observation of the presence of Ehrlichia sp. and Anaplasma sp. was used as a positivity criterion. 5,015 animals were treated during this period and 691 (13.78%) of them were positive for the presence of these hemoparasites. Among these animals, 9.23% (463/5015) had morulae from Ehrlichia sp., and 4.37% (219/5015) had morulae typical of Anaplasma sp., while 0.18% (9/5015) had morulae of both species. Females represented 57.89% (400/691) of positive animals and males 42.11% (291/691). 35.02% (242/691) of the animals were less than one year old, 54.13% (374/691) were between 1 and 8 years old, and 10.85% (75/691) were over 8 years old. Females, less than one year old and co-infected, showed the most significant changes in hematological, hematocrit (29%), red blood cell count (3 million / mm3), platelet count (40,000 / mm3) and total blood count leukocytes (16,000 thousand / mm3), showing anemia and thrombocytopenia. Animals infected with Anaplasma sp. and Ehrlichia sp., at the same time, presented more intense thrombocytopenia, evidenced in males between 1 and 8 years old (30,000 / mm3). Infections by Anaplasma sp. caused a severe decrease in the mean red blood cell count, evidenced in males (3 million / mm3) and females under 1 year (3.79 million / mm3).

En Brasil, las infecciones por hemoparásitos representan una de las principales causas de búsqueda de asistencia veterinaria en perros. Infecciones por bacterias del orden Rickettsiales que incluyen Ehrlichia sp.y Anaplasma sp.tienen una gran participación en estas búsquedas. El objetivo de este estudio fue dilucidar los aspectos epidemiológicos regionales y hematológicos de las infecciones por Anaplasma sp.y Ehrlichia sp., muchas veces asintomáticas, en perros del municipio de Jataí-GO. Los datos de este estudio se basaron en muestras de sangre de perros tratados en el Hospital Veterinario de la Universidad Federal de Jataí, en Jataí, Goiás, entre los años 2005 y 2017. Observación microscópica de la presencia de Ehrlichia sp. y Anaplasma sp. se utilizó como criterio de positividad. Durante este período se trataron 5.015 animales y 691 (13,78%) de ellos resultaron positivos para la presencia de estos hemoparásitos. Entre estos animales, 9,23% (463/5015) tenía mórulas de Ehrlichia sp., y 4,37% (219/5015) tenían mórulas típicas de Anaplasma sp., mientras que 0,18% (9/5015) tenían mórulas de ambas especies. Las hembras representaron 57,89% (400/691) de los animales positivos y los machos 42,11% (291/691). 35,02% (242/691) de los animales tenían menos de 1 año, 54,13% (374/691) tenían entre 1 y 8 años y 10,85% (75/691) tenían más de 8 años. Las hembras, menores de un año y coinfectadas, mostraron los cambios más significativos en hematología, hematocrito (29%), recuento de glóbulos rojos (3 millones/mm3), recuento de plaquetas (40.000/mm3) y recuento sanguíneo total leucocitos (16.000 mil/mm3), mostrando anemia y trombocitopenia. Los animales infectados por Anaplasma sp. y Ehrlichia sp., al mismo tiempo, presentaron trombocitopenia más intensa, evidenciada en machos entre 1 y 8 años (30.000 /mm3). Infecciones por Anaplasma sp. causó una disminución severa en el recuento promedio de glóbulos rojos, evidenciada en machos (3 millones/mm3) y hembras menores de 1 año (3.79 millones/mm3).

Aspectos epidemiológicos e hematológicos de cães infectados com Ehrlichia sp e Anaplasma sp em Jataí - GO, Brasil / Epidemiological and hematological aspects of dogs with Ehrlichia sp and Anaplasma sp in Jataí - GO / Aspectos epidemiológicos y hematológicos de perros con Ehrlichia sp y Anaplasma sp en Jataí - GO

No Brasil, as infecções por hemoparasitas representam uma das principais causas de busca por assistência veterinária em cães. Infecções por bactérias da ordem Rickettsiales que incluem Ehrlichia sp. e Anaplasma sp.tem uma grande participação nessas buscas. Foi objetivo deste estudo elucidar aspectos epidemiológicos regionais e hematológicos de infecções por Anaplasma sp.e Ehrlichia sp, muitas vezes assintomáticas, em cães no município de Jataí-GO. Os dados deste estudo foram baseados em amostras de sangue de cães atendidos no Hospital Veterinário da Universidade Federal de Jataí, em Jataí, Goiás, entre os anos de 2005 e 2017. A observação microscópica da presença de mórulas de Ehrlichia sp. e Anaplasma sp. foi utilizada como critério de positividade. 5.015 animais foram atendidos nesse período e 691 (13,78%) deles foram positivos para a presença destes hemoparasitas. Entre esses animais, 9,23% (463/5015) possuíam mórulas de Ehrlichia sp., e 4,37% (219/5015) apresentavam mórulas típicas de Anaplasma sp.,enquanto 0,18% (9/5015) possuíam mórula de ambas as espécies. As fêmeas representaram 57,89% (400/691) dos animais positivos e os machos 42,11% (291/691). 35,02% (242/691) dos animais tinham menos de 1 ano de vida, 54,13% (374/691) tinham entre 1 e 8 anos, e 10,85% (75/691) tinham mais de 8 anos. As fêmeas, com menos de um ano de idade e coinfectadas, apresentaram as alterações mais significativas nos aspectos hematológicos, hematócrito (29%), contagem de hemácias (3 milhões/mm3), contagem de plaquetas (40.000/mm3) e contagem total de leucócitos (16.000 mil/mm3), evidenciando anemia e trombocitopenia. Animais infectados com Anaplasma sp.e Ehrlichia sp.,ao mesmo tempo, apresentaram trombocitopenia mais intensa, evidenciada em machos entre 1 e 8 anos (30.000/mm3). As infecções por Anaplasma sp. causaram uma diminuição severa na média da contagem de glóbulos vermelhos, evidenciada em machos (3 milhões/mm3) e fêmeas com menos de 1 ano (3,79 milhões/mm3).(AU)

In Brazil, infections by hemoparasites represent one of the main causes of seeking veterinary assistance in dogs. Infections with bacteria of the order Rickettsiales that include Ehrlichia sp. and Anaplasma sp. has a great participation in these assistances. The objective of this study was to elucidate regional epidemiologic and hematologic aspects of infections by Anaplasma sp. and Ehrlichia sp.,often asymptomatic, in dogs in the municipality of Jataí-GO. The data in this study were based on blood samples from dogs treated at the Veterinary Hospital of the Federal University of Jataí, in Jataí, Goiás, between the years 2005 and 2017. Microscopic observation of the presence of Ehrlichia sp. and Anaplasma sp. was used as a positivity criterion. 5,015 animals were treated during this period and 691 (13.78%) of them were positive for the presence of these hemoparasites. Among these animals, 9.23% (463/5015) had morulae from Ehrlichia sp., and 4.37% (219/5015) had morulae typical of Anaplasma sp., while 0.18% (9/5015) had morulae of both species. Females represented 57.89% (400/691) of positive animals and males 42.11% (291/691). 35.02% (242/691) of the animals were less than one year old, 54.13% (374/691) were between 1 and 8 years old, and 10.85% (75/691) were over 8 years old. Females, less than one year old and co-infected, showed the most significant changes in hematological, hematocrit (29%), red blood cell count (3 million / mm3), platelet count (40,000 / mm3) and total blood count leukocytes (16,000 thousand / mm3), showing anemia and thrombocytopenia. Animals infected with Anaplasma sp. and Ehrlichia sp., at the same time, presented more intense thrombocytopenia, evidenced in males between 1 and 8 years old (30,000 / mm3). Infections by Anaplasma sp. caused a severe decrease in the mean red blood cell count, evidenced in males (3 million / mm3) and females under 1 year (3.79 million / mm3).(AU)

En Brasil, las infecciones por hemoparásitos representan una de las principales causas de búsqueda de asistencia veterinaria en perros. Infecciones por bacterias del orden Rickettsiales que incluyen Ehrlichia sp.y Anaplasma sp.tienen una gran participación en estas búsquedas. El objetivo de este estudio fue dilucidar los aspectos epidemiológicos regionales y hematológicos de las infecciones por Anaplasma sp.y Ehrlichia sp., muchas veces asintomáticas, en perros del municipio de Jataí-GO. Los datos de este estudio se basaron en muestras de sangre de perros tratados en el Hospital Veterinario de la Universidad Federal de Jataí, en Jataí, Goiás, entre los años 2005 y 2017. Observación microscópica de la presencia de Ehrlichia sp. y Anaplasma sp. se utilizó como criterio de positividad. Durante este período se trataron 5.015 animales y 691 (13,78%) de ellos resultaron positivos para la presencia de estos hemoparásitos. Entre estos animales, 9,23% (463/5015) tenía mórulas de Ehrlichia sp., y 4,37% (219/5015) tenían mórulas típicas de Anaplasma sp., mientras que 0,18% (9/5015) tenían mórulas de ambas especies. Las hembras representaron 57,89% (400/691) de los animales positivos y los machos 42,11% (291/691). 35,02% (242/691) de los animales tenían menos de 1 año, 54,13% (374/691) tenían entre 1 y 8 años y 10,85% (75/691) tenían más de 8 años. Las hembras, menores de un año y coinfectadas, mostraron los cambios más significativos en hematología, hematocrito (29%), recuento de glóbulos rojos (3 millones/mm3), recuento de plaquetas (40.000/mm3) y recuento sanguíneo total leucocitos (16.000 mil/mm3), mostrando anemia y trombocitopenia. Los animales infectados por Anaplasma sp. y Ehrlichia sp., al mismo tiempo, presentaron trombocitopenia más intensa, evidenciada en machos entre 1 y 8 años (30.000 /mm3). Infecciones por Anaplasma sp. causó una disminución severa en el recuento promedio de glóbulos rojos, evidenciada en machos (3 millones/mm3) y hembras menores de 1 año (3.79 millones/mm3).(AU)

Successful treatment and management of canine ehrlichiosis-leishmaniosis-heartworm comorbidity

Background: Canine vector borne diseases (CVBD) are common in tropical countries where the climate favors arthropods abundance. Comorbidity with one or more CVBD are reported as clinical complication and worsen prognostic. Canine visceral leishmaniosis (CanL) is an endemic zoonotic disease in Brazil caused by Leishmania infantum, with several restrictions to canine treatment and suggestion of reservoirs euthanasia for disease control. Heartworm (HW) is a helminthic disease caused by Dirofilaria immitis infection in dogs. It is a chronic heart disease, which can lead to death by congestive heart failure. Canine ehrlichiosis (CE) is caused by Ehrlichia canis bacterial infection with a zoonotic potential and fatal to dogs in acute and chronic presentations. Exposed the above, this study aims to describe a successful treatment and management of a dog with CanL, CE, and HW comorbidity. Case: A 3-year-old male uncastrated black Labrador dog, weighing 35 kg, was admitted to the veterinary clinic due to immunochromatographic CanL positive test performed by municipal zoonosis control center active surveillance. Clinical exam showed a mild shedding, intermittent eye white/yellow discharge and popliteal lymph nodes enlargement. After positive for CanL, veterinary requested more laboratorial exams. IFAT and ELISA were positive for CanL, blood smear showed presence of microfilaria, and bone marrow cytology showed Ehrlichia spp. morulae and microfilaria. Initial treatment prescribed was oral doxycycline, omeprazole, ranitidine, and domperidone for 30 days, and allopurinol and ivermectin until further recommendation. Additionally, repellent collar, repellent spray and vitamin supplement was indicated. After first month, marbofloxacin for 30 days and three doses of immunostimulant drug were administrated. After three months of treatment, dog still positive for heartworm, ehrlichiosis, and CanL. Doxycycline protocol was repeated. Dog became consistently negative for all pathogens one year later with persistent thrombocytopenia but without clinical signs, ergo allopurinol and ivermectin were discontinued. After 4 years of follow up, the animal had an acute pancreatitis and died, with unremarkable total blood count and negative for all pathogens. Discussion: CVBD coinfections are commonly reported as worsen prognostic in endemic regions. The pathogens reported here share a host immunomodulation competence. L. infantum and Ehrlichia spp. downregulates Th1 response, whereas D. immitis increase as Th2 profile. The therapeutic protocol was iniciated by staging CanL. Since the patient had clinical signs, allopurinol was prescribed as a well-established drug for CanL. Marbofloxacin was added due to its high safety drug in clinical improvement of infected dogs with and without renal disease and in vitro effectiveness against L. infantum. Domperidone was used to promote Th1 cytokine profile as INF-γ, IL-2, IL-12, and TNF-α. We used an immunostimulant protocol to favor polarization to the Th1 profile comprised by 30 days of domperidone protocol followed by a vaccine and an immunomodulator. Doxycycline was used successfully for Ehrlichia spp. and HE clearance after 2 treatment courses and 1 year of ivermectin every 15 days. The animal presented intermittent coughing episodes on the first treatment course, but no medical intervention was needed besides exercise restriction. Our report shows the successful management of one dog with CanL, CE and HE comorbidity. This success was possible due to early detection and good therapeutic choice.(AU)

Produção de embriões partenogenéticos de cutias usando Cloreto de Estrôncio e Citocalasina B / Production of parthenogenetic embryos from agoutis using strontium chloride and cytochalasin B

The study aimed to evaluate the effect of strontium chloride (SrCl2) with cytochalasin B (CB) on the activation of agouti oocytes matured in vitro for embryo production. Thus, ovaries were used for oocyte recovery by slicing. Subsequently, viable oocytes were destined for in vitro maturation (IVM) and after 24 h evaluated for the expansion and viability of the cumulus cells and presence of the first polar body (1PB). After IVM, oocytes were activated with a combination of 10 mM SrCl2 and 5 μg/mL CB for 6 h and evaluated for embryo development kinetics. Hence, 93.3% of cumulus cell expansion was observed, with 91.2% viability and 37.3% of oocytes with the presence of 1PB. Regarding embryonic development, 43.2% (19/44) of cleaved structures and 6.8% (3/44) of morulae were observed in relation to the number of oocytes and 18.8% (3/16) morulae in relation to the number of cleaved structures. Thus, the combination of SrCl2 with CB promoted the activation of oocytes matured in vitro from agouti resulting in morulae. Finally, with this study, fundamental steps for in vitro conservation through reproductive biotechniques were developed in agoutis.

Produção de embriões partenogenéticos de cutias usando Cloreto de Estrôncio e Citocalasina B / Production of parthenogenetic embryos from agoutis using strontium chloride and cytochalasin B

The study aimed to evaluate the effect of strontium chloride (SrCl2) with cytochalasin B (CB) on the activation of agouti oocytes matured in vitro for embryo production. Thus, ovaries were used for oocyte recovery by slicing. Subsequently, viable oocytes were destined for in vitro maturation (IVM) and after 24 h evaluated for the expansion and viability of the cumulus cells and presence of the first polar body (1PB). After IVM, oocytes were activated with a combination of 10 mM SrCl2 and 5 μg/mL CB for 6 h and evaluated for embryo development kinetics. Hence, 93.3% of cumulus cell expansion was observed, with 91.2% viability and 37.3% of oocytes with the presence of 1PB. Regarding embryonic development, 43.2% (19/44) of cleaved structures and 6.8% (3/44) of morulae were observed in relation to the number of oocytes and 18.8% (3/16) morulae in relation to the number of cleaved structures. Thus, the combination of SrCl2 with CB promoted the activation of oocytes matured in vitro from agouti resulting in morulae. Finally, with this study, fundamental steps for in vitro conservation through reproductive biotechniques were developed in agoutis.(AU)

Vitrified embryo transfer in Merino sheep under extensive conditions

The aim was to evaluate pregnancy success after transfer of embryos vitrified in micropipette tips in Merino sheep under extensive conditions. A second objective was to evaluate the influence of embryo stage in such pregnancy rate. One hundred and twenty-seven embryos were rewarmed and transferred into recipient ewes. On rewarming, the embryos were placed into three-step cryoprotectant dilutions. Finally, prior to transfer to recipient females, embryos were maintained in Basic Medium for 5 min at 25ºC and were reevaluated by morphological criteria; all degenerated embryos were eliminated. Recipient ewes (n = 150) were treated for estrus with sponges placed for 14 days and 300 IU of eCG. At embryo transfer, three experimental groups were defined: morulae transferred on Day 7, blastocysts transferred on Day 7 and blastocysts transferred on Day 8 after sponge removal. In all groups, semi-laparoscopic transfer of one rewarmed embryo per recipient was performed. Pregnancy was diagnosed by ultrasonography on day 28 after embryo transfer. The embryo selection rate after rewarming was higher for blastocysts (89.3% - 67/75) compared to morulae (65.9% - 60/91) (P < 0.05). Pregnancy diagnosis showed a 38.3% (23/60) of success after morula transfer on Day 7 post progestagen removal. The day of transfer showed a significant influence on pregnancy rate after blastocyst transfer (Day 8, 55.9% - 19/34 vs Day 7, 21.2% - 7/33) (P < 0.05). Blastocysts transfer on Day 8 showed the highest global efficiency (pregnancies/total embryos after rewarming) (47.5% - 19/40) (P < 0.05). In conclusion, reproductive efficiency obtained by vitrified embryo transfer allows its recommendation for embryo transfer programs under extensive conditions. The importance of considering the synchrony between the embryo age and the recipient uterus stage is emphasized.

Vitrified embryo transfer in Merino sheep under extensive conditions

The aim was to evaluate pregnancy success after transfer of embryos vitrified in micropipette tips in Merino sheep under extensive conditions. A second objective was to evaluate the influence of embryo stage in such pregnancy rate. One hundred and twenty-seven embryos were rewarmed and transferred into recipient ewes. On rewarming, the embryos were placed into three-step cryoprotectant dilutions. Finally, prior to transfer to recipient females, embryos were maintained in Basic Medium for 5 min at 25ºC and were reevaluated by morphological criteria; all degenerated embryos were eliminated. Recipient ewes (n = 150) were treated for estrus with sponges placed for 14 days and 300 IU of eCG. At embryo transfer, three experimental groups were defined: morulae transferred on Day 7, blastocysts transferred on Day 7 and blastocysts transferred on Day 8 after sponge removal. In all groups, semi-laparoscopic transfer of one rewarmed embryo per recipient was performed. Pregnancy was diagnosed by ultrasonography on day 28 after embryo transfer. The embryo selection rate after rewarming was higher for blastocysts (89.3% - 67/75) compared to morulae (65.9% - 60/91) (P < 0.05). Pregnancy diagnosis showed a 38.3% (23/60) of success after morula transfer on Day 7 post progestagen removal. The day of transfer showed a significant influence on pregnancy rate after blastocyst transfer (Day 8, 55.9% - 19/34 vs Day 7, 21.2% - 7/33) (P < 0.05). Blastocysts transfer on Day 8 showed the highest global efficiency (pregnancies/total embryos after rewarming) (47.5% - 19/40) (P < 0.05). In conclusion, reproductive efficiency obtained by vitrified embryo transfer allows its recommendation for embryo transfer programs under extensive conditions. The importance of considering the synchrony between the embryo age and the recipient uterus stage is emphasized.(AU)

Reproductive physiology of the heat-stressed dairy cow: implications for fertility and assisted reproduction

Heat stress causes a large decline in pregnancy success per insemination during warm times of the year. Improvements in fertility are possible by exploiting knowledge about how heat stress affects the reproductive process. The oocyte can be damaged by heat stress at the earliest stages of folliculogenesis and remains sensitive to heat stress in the peri-ovulatory period. Changes in oocyte quality due to heat stress are the result of altered patterns of folliculogenesis and, possibly, direct effects of elevated body temperature on the oocyte. While adverse effects of elevated temperature on the oocyte have been observed in vitro, local cooling of the ovary and protective effects of follicular fluid may limit these actions in vivo. Heat stress can also compromise fertilization rate. The first seven days of embryonic development are very susceptible to disruption by heat stress. During these seven days, the embryo undergoes a rapid change in sensitivity to heat stress from being very sensitive (2- to 4-cell stage) to largely resistant (by the morulae stage). Direct actions of elevated temperature on the embryo are likely to be an important mechanism for reduction in embryonic survival caused by heat stress. An effective way to avoid effects of heat stress on the oocyte, fertilization, and early embryo is to bypass the effects through embryo transfer because embryos are typically transferred into females after acquisition of thermal resistance. There may be some opportunity to mitigate effects of heat stress by feeding antioxidants or regulating the endocrine environment of the cow but neither approach has been reduced to practice. The best long-term solution to the problem of heat stress may be to increase genetic resistance of cows to heat stress. Thermotolerance genes exist within dairy breeds and additional genes can be introgressed from other breeds by traditional means or gene editing.

Reproductive physiology of the heat-stressed dairy cow: implications for fertility and assisted reproduction

Heat stress causes a large decline in pregnancy success per insemination during warm times of the year. Improvements in fertility are possible by exploiting knowledge about how heat stress affects the reproductive process. The oocyte can be damaged by heat stress at the earliest stages of folliculogenesis and remains sensitive to heat stress in the peri-ovulatory period. Changes in oocyte quality due to heat stress are the result of altered patterns of folliculogenesis and, possibly, direct effects of elevated body temperature on the oocyte. While adverse effects of elevated temperature on the oocyte have been observed in vitro, local cooling of the ovary and protective effects of follicular fluid may limit these actions in vivo. Heat stress can also compromise fertilization rate. The first seven days of embryonic development are very susceptible to disruption by heat stress. During these seven days, the embryo undergoes a rapid change in sensitivity to heat stress from being very sensitive (2- to 4-cell stage) to largely resistant (by the morulae stage). Direct actions of elevated temperature on the embryo are likely to be an important mechanism for reduction in embryonic survival caused by heat stress. An effective way to avoid effects of heat stress on the oocyte, fertilization, and early embryo is to bypass the effects through embryo transfer because embryos are typically transferred into females after acquisition of thermal resistance. There may be some opportunity to mitigate effects of heat stress by feeding antioxidants or regulating the endocrine environment of the cow but neither approach has been reduced to practice. The best long-term solution to the problem of heat stress may be to increase genetic resistance of cows to heat stress. Thermotolerance genes exist within dairy breeds and additional genes can be introgressed from other breeds by traditional means or gene editing.(AU)

Supplementing alfa-Linolenic acid in the in vitro maturation media improves nuclear maturation rate of oocytes and early embryonic development in the Nili Ravi buffalo

The present study was conducted to investigate the effect of omega-3 poly unsaturated fatty acid (PUFA), α-linolenic acid (ALA; 18:3 n-3) on the in vitro maturation (IVM) of buffalo oocytes and subsequent embryonic development. Buffalo cumulus oocyte complexes (COCs; n = 2282) were in vitro matured in TCM-199 (0.6% fatty acid free bovine serum albumin, 0.02 Units/ml FSH, 1 µg/ml 17-β-estradiol, 10 µg/ml epidermal growth factor, 50 µg/ml gentamicin) supplemented with 0 (control), 25, 50, 100, 150 or 300 µm ALA under an atmosphere of 5% CO2 in air at 38.5ºC for 22-24 h. The matured oocytes were then fertilized in Tyrode’s Albumin Lactate Pyruvate (TALP) medium and cultured in synthetic oviductal fluid (SOF) medium. Concentrations up to 100 μm ALA improves (P ≤ 0.05) the cumulus expansion compared to control. Higher percentage of oocytes reaching MII stage was observed at 50 μm and 100 μm of ALA compared to control (P ≤ 0.05). Concentrations of 150 and 300 µm ALA were detrimental both for cumulus expansion and nuclear maturation rate of buffalo oocytes. Moreover, supplementation with 100 μm ALA improved (P ≤ 0.05) cleavage rate compared to control and treatment with 50 and 100 μm ALA yielded significantly higher morulae compared to control. The results of present study indicate that the supplementation with 100 μm ALA to the IVM medium improves nuclear maturation rate of buffalo oocytes and subsequent early embryonic development.

Supplementing alfa-Linolenic acid in the in vitro maturation media improves nuclear maturation rate of oocytes and early embryonic development in the Nili Ravi buffalo

The present study was conducted to investigate the effect of omega-3 poly unsaturated fatty acid (PUFA), α-linolenic acid (ALA; 18:3 n-3) on the in vitro maturation (IVM) of buffalo oocytes and subsequent embryonic development. Buffalo cumulus oocyte complexes (COCs; n = 2282) were in vitro matured in TCM-199 (0.6% fatty acid free bovine serum albumin, 0.02 Units/ml FSH, 1 µg/ml 17-β-estradiol, 10 µg/ml epidermal growth factor, 50 µg/ml gentamicin) supplemented with 0 (control), 25, 50, 100, 150 or 300 µm ALA under an atmosphere of 5% CO2 in air at 38.5ºC for 22-24 h. The matured oocytes were then fertilized in Tyrodes Albumin Lactate Pyruvate (TALP) medium and cultured in synthetic oviductal fluid (SOF) medium. Concentrations up to 100 μm ALA improves (P ≤ 0.05) the cumulus expansion compared to control. Higher percentage of oocytes reaching MII stage was observed at 50 μm and 100 μm of ALA compared to control (P ≤ 0.05). Concentrations of 150 and 300 µm ALA were detrimental both for cumulus expansion and nuclear maturation rate of buffalo oocytes. Moreover, supplementation with 100 μm ALA improved (P ≤ 0.05) cleavage rate compared to control and treatment with 50 and 100 μm ALA yielded significantly higher morulae compared to control. The results of present study indicate that the supplementation with 100 μm ALA to the IVM medium improves nuclear maturation rate of buffalo oocytes and subsequent early embryonic development.(AU)

Ocorrência de infecção concomitante natural por Ehrlichia sp., Anaplasma sp. e Babesia sp. em cão assistido no Hospital Veterinário da Universidade Federal de Uberlândia / Occurrence of concomitant natural infection by Ehrlichia sp., Anaplasma sp. and Babesia sp. in a dog assisted at the Hospital of the Veterinary University of Uberlândia

Foi atendido no ambulatório do Hospital Veterinário da Universidade Federal de Uberlândia um canino sem raça definida de quatro anos de idade, domiciliado. Na anamnese o proprietário relatou que o cão estava apático, inapetente, com prurido e dificuldade respiratória. Durante o exame clínico específico observou-se hipertrofia dos linfonodos poplíteos, esplenomegalia, além de uma discreta hipertermia. O hemograma revelou trombocitopenia e leucopenia com eosinopenia e monocitopenia. No esfregaço sanguíneo periférico foram encontradas mórulas de Ehrlichiasp. em monócitos, mórulas de Anaplasma sp. em plaquetas e inclusões intraeritrocitárias de Babesia sp. O animal foi tratado com doxiciclina e imizol, tendo seu retorno após quinze dias do início do tratamento, e ao realizar novos exames hematológicos os resultados se mostraram normais sem nenhum achado na pesquisa de hemoparasitos, o que indica o sucesso do tratamento proposto. (AU)

It was treated at the Hospital of the Veterinary University of Uberlândia a mixed breed dog with four years old, domiciled. In the interview the owner reported that the canine was lethargic, inapetente with itching and difficulty breathing. During the specific clinical examination there was hypertrophy of popliteal lymph nodes, splenomegaly, and a slight hyperthermia. The blood count revealed a thrombocytopenia, leukopenia, and with eosinopeniamonocytopenia. In peripheral blood smear was found morulae of Ehrlichia sp. into monocytes, morulaof Anaplasma sp. into platelets and inclusions intraerytrocytic Babesia sp. The animal was treated with doxycycline and imizol and hashis return after fifteen days of starting treatment, and after making new blood tests the results were normal with no finding in hemoparasite research, which indicates the success of the proposed treatment. (AU)

Erliquiose canina - relato de caso / Canine ehrlichiosis - report case

The ehrlichiosis is a disease transmitted through blood meal of Rhipicephalus sanguineus tick bites infected with the bacteria Ehrlichia spp. acquired from another infected dog. Signs include apathy, anorexia, fever may reach severe thrombocytopenia and marrow aplasia. The objective of this article is to report a case of ehrlichiosis in a dog. The diagnosis is made by clinical history with the presence of ectoparasites, compatible clinical signs and blood count, which may present anemia, thrombocytopenia, pancytopenia and morulae of Erlichia spp. For treatment, doxycycline is the drug of choice and should be used for 28 days.

Erliquiose canina - relato de caso / Canine ehrlichiosis - report case

The ehrlichiosis is a disease transmitted through blood meal of Rhipicephalus sanguineus tick bites infected with the bacteria Ehrlichia spp. acquired from another infected dog. Signs include apathy, anorexia, fever may reach severe thrombocytopenia and marrow aplasia. The objective of this article is to report a case of ehrlichiosis in a dog. The diagnosis is made by clinical history with the presence of ectoparasites, compatible clinical signs and blood count, which may present anemia, thrombocytopenia, pancytopenia and morulae of Erlichia spp. For treatment, doxycycline is the drug of choice and should be used for 28 days.(AU)

Ocorrência de infecção concomitante natural por Ehrlichia sp., Anaplasma sp. e Babesia sp. em cão assistido no Hospital Veterinário da Universidade Federal de Uberlândia / Occurrence of concomitant natural infection by Ehrlichia sp., Anaplasma sp. and Babesia sp. in a dog assisted at the Hospital of the Veterinary University of Uberlândia

Foi atendido no ambulatório do Hospital Veterinário da Universidade Federal de Uberlândia um canino sem raça definida de quatro anos de idade, domiciliado. Na anamnese o proprietário relatou que o cão estava apático, inapetente, com prurido e dificuldade respiratória. Durante o exame clínico específico observou-se hipertrofia dos linfonodos poplíteos, esplenomegalia, além de uma discreta hipertermia. O hemograma revelou trombocitopenia e leucopenia com eosinopenia e monocitopenia. No esfregaço sanguíneo periférico foram encontradas mórulas de Ehrlichiasp. em monócitos, mórulas de Anaplasma sp. em plaquetas e inclusões intraeritrocitárias de Babesia sp. O animal foi tratado com doxiciclina e imizol, tendo seu retorno após quinze dias do início do tratamento, e ao realizar novos exames hematológicos os resultados se mostraram normais sem nenhum achado na pesquisa de hemoparasitos, o que indica o sucesso do tratamento proposto.

It was treated at the Hospital of the Veterinary University of Uberlândia a mixed breed dog with four years old, domiciled. In the interview the owner reported that the canine was lethargic, inapetente with itching and difficulty breathing. During the specific clinical examination there was hypertrophy of popliteal lymph nodes, splenomegaly, and a slight hyperthermia. The blood count revealed a thrombocytopenia, leukopenia, and with eosinopeniamonocytopenia. In peripheral blood smear was found morulae of Ehrlichia sp. into monocytes, morulaof Anaplasma sp. into platelets and inclusions intraerytrocytic Babesia sp. The animal was treated with doxycycline and imizol and hashis return after fifteen days of starting treatment, and after making new blood tests the results were normal with no finding in hemoparasite research, which indicates the success of the proposed treatment.

Survey of Ehrlichia canis, Babesia spp. and Hepatozoon spp. in dogs from a semiarid region of Brazil

This study assessed the occurrence of Ehrlichia spp., Babesia spp. and Hepatozoon spp. infections in 100 tick-harboring dogs from a semiarid region of the State of Paraíba, Northeastern Brazil. Blood samples and ticks were collected from the animals, and a questionnaire was submitted to dog owners to obtain general data. Blood samples were used to perform hemogram, direct blood smear and immunological and molecular hemoparasite detection. The 1,151 ticks collected were identified as Rhipicephalus sanguineus; direct smears revealed E. canis-like morulae in the monocytes of 4% (4/100) of the non-vaccinated female dogs, and 34% and 25% of the dogs tested positive for Ehrlichia canis by indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR), respectively. Blood smear examination revealed Babesia-suggestive merozoites in the erythrocytes of 2% (2/100) of the animals. Babesia vogeli was detected by PCR in ten animals (10%) and was correlated with young age (p = 0.007) and thrombocytopenia (p = 0.01). None of the animals showed Hepatozoon spp. positivity. These results indicate that E. canis is the main tick-borne canine pathogen in the study area and provide the first report of B. vogeli infection in dogs from Paraiba State.

O presente estudo avaliou a ocorrência de infecção por Ehrlichia spp., Babesia spp. e Hepatozoon spp. em 100 cães, infestados por carrapatos, oriundos de uma região semiárida do Estado da Paraíba, Nordeste do Brasil. Amostras de sangue e de carrapatos foram coletadas dos animais, e um questionário foi submetido aos proprietários dos cães para obter dados gerais. As amostras de sangue foram utilizadas para realização de hemograma, esfregaço sanguíneo e detecção molecular dos hemoparasitos. Os 1.151 carrapatos coletados foram identificados como Rhipicephalus sanguineus; os esfregaços sanguíneos revelaram mórulas sugestivas de E. canis em 4% (4/100) de cães fêmeas não vacinadas, e 34% e 25% dos cães foram positivos para Ehrlichia canis pela imunofluorescência indireta (IFI) e reação em cadeia pela polimerase (PCR), respectivamente. Os esfregaços sanguíneos revelaram merozoítas sugestivas de Babesia em eritrócitos de 2% (2/100) dos animais. Babesia vogeli foi detectada por PCR em dez animais (10%) e foi correlacionada com a idade jovem (p=0,007) e trombocitopenia (p=0,01). Nenhum dos animais apresentou positividade para Hepatozoon spp. Esses resultados indicam que E. canis é o principal patógeno canino transmitido por carrapato, na área estudada, e fornece o primeiro relato de infecção por B. vogeli em cães do Estado da Paraíba.

DIAGNÓSTICO DE Ehrlichia spp. EM CÃES ASSINTOMÁTICOS

A Ehrlichia canis é um importante agente infeccioso em cães, de difícil controle devido a distribuição do seu vetor, o Rhipicephalus sanguineus. Sendo transmitida de um cão para outro pelo repasto sanguíneo do carrapato vetor, que pela hematofagia ingere o agente de um cão infectado contendo mórulas nas células parasitadas. Durante o repasto sanguíneo, os componentes da saliva do carrapato atuam na resposta imune local do hospedeiro, diminuindo a resposta do tipo Th1. Essa supressão favorece a resposta do tipo Th2, possibilitando que a infecção se instale. Desse modo o animal gera alterações hematológicas e bioquímicas inespecíficas dificultando o diagnóstico, que muitas vezes é feito de forma presuntiva. Induzindo ao uso de medicação desnecessária podendo gerar complicações na eficácia do tratamento devido ao favorecimento de resistência a antibióticos. É importante esclarecer e estudar sobre os meios de diagnósticos e as associações necessárias, para evitar tratamentos desnecessários, especialmente em casos assintomáticos. O diagnóstico definitivo requer o uso de técnicas mais sensíveis como técnicas moleculares e não somente considerar o hemograma completo e testes sorológicos qualitativos. Visto que a disseminação de casos de erliquiose entre a população canina e as limitações das técnicas convencionais de diagnóstico, identificação e diferenciação de microrganismos podem promover equívocos no diagnóstico de doenças infecciosas, pretende-se explorar outros meios de diagnósticos disponíveis em cães assintomáticos com alterações hematológicas, visando esclarecer e conscientizar sobre a forma como os tratamentos contra hemoparasitoses estão sendo administrados.

Ehrlichia canis is an important infectious agent in dogs, difficult to control due to the distribution of its vector, Rhipicephalus sanguineus. Being transmitted from one dog to another by the blood meal of the vector tick, which by hematophagy ingests the agent of an infected dog containing morulae in the parasitized cells. During blood meal, the components of the tick's saliva act on the host's local immune response, decreasing the Th1-type response. This suppression favors the Th2-type response, allowing the infection to settle. In this way, the animal generates nonspecific hematological and biochemical changes, making the diagnosis difficult, which is often presumptively made. Inducing the use of unnecessary medication, which can lead to complications in the effectiveness of the treatment due to favoring resistance to antibiotics. It is important to clarify and study the means of diagnosis and the necessary associations, to avoid unnecessary treatments, especially in asymptomatic cases. The definitive diagnosis requires the use of more sensitive techniques such as molecular techniques and not only considering the complete blood count and qualitative serological tests. Since the spread of cases of ehrlichiosis among the canine population and the limitations of conventional techniques for the diagnosis, identification and differentiation of microorganisms can promote misunderstandings in the diagnosis of infectious diseases, it is intended to explore other means of diagnosis available in asymptomatic dogs with hematological changes, aiming to clarify and raise awareness about how treatments against hemoparasitosis are being administered.

Serum TNF- and IL-10 in Ehrlichia spp. naturally infected dogs

Background: Canine ehrlichiosis (CE) is a disease caused by Ehrlichia canis, transmitted by Rhipicephalus sanguineus and dogs can present variable clinical manifestations, ranging from subclinical, acute to chronic illness. This pathogen preferentially infects leukocytes. In Brazil illness is described nationwide with the seropositivity varying from 4.8% to over 50%. The control of Ehrlichia spp. infection was related to cellular immune response mediated by Th1 while the CE acute phase is related to humoral immune response mediated by Th2, with high production of antibodies. Th1 and Th2 balance can be measured by cytokine profile. TNF-α is associated with inflammatory responses while IL-10 inhibits mechanisms which prevent tissue damage caused by the intense inflammatory response. The aim of this study was to evaluate the TNF-α and IL-10 serum levels in naturally infected dogs by Ehrlichia spp.Material, Methods & Results: Twenty five dogs, both sexes, one to eight years old, weighing 5 to 40 kg, undefined breed were divided in two groups: Control (CT, n = 14) and naturally infected by Ehrlichia spp. dogs (EH, n = 11). Naturally infected dogs were from Zoonosis Control Center. Ehrlichia spp. morulae was visualized in bone marrow aspirates. Blood samples were collected for hematological analysis and serum dosages of TNF-α and IL-10 (commercial kits by sandwich ELISA). The frequency of clinical signs was expressed as percentage. T test, Spearman correlation and linear regression were used when necessary (P ≤ 0.05). Alopecia (45%), generalized lymphadenopathy (36%), onychogryphosis (27%) and conjunctivitis (27%) were the most frequent clinical signs in the EH group. No difference was found between studied groups in hematological and biochemical parameters. TNF-α concentration was 43.3 ± 16.9 pg/mL in CT and 41.1 ± 3.8 pg/mL in EH, while IL-10 concentration was 20.2 ± 8.4 pg/mL in CT and 21.0 ± 5.1 pg/mL in EH.[...]

Effects of exposure to halothane, isoflurane, and sevoflurane on embryo viability and gestation in female mice / Efeitos da exposição ao halotano, isofluorano, e sevofluorano na viabilidade embrionária e gestação em fêmeas de camundongos

Although the negative effects of inhalation anaesthetics on fertility have been known for a while, the stages during the reproductive cycle at which these effects occur and the mechanisms of action are largely unknown. This study aimed to evaluate the effects of acute exposure of female mice to halothane, isoflurane, and sevoflurane prior to mating. BALB/c female mice (n=160) were allocated in groups of 20 to halothane (HG), isoflurane (IG), sevoflurane (SG), and oxygen/sham (SH) treatment groups and their respective control groups (CGs). The mice were exposed to 1 minimum alveolar concentration (MAC) of the corresponding anaesthetic or oxygen for 4 h/day over 5 consecutive days. Two days after exposure, females were mated with males (ratio 2:1 female/male) for five consecutive days. Every morning, females were checked for the presence of vaginal plugs. Half of the females that exhibited plugs were euthanised 72 h later for embryo evaluation. The remaining females were euthanised on the 14th day of pregnancy for foetal evaluation. A low number of morulae and total embryos (morulae + blastocysts) were observed in the HG (P 0.05). The number of implantations was lower in the HG (6.0) compared with the IG (11.8) and SG (12.4) (P 0.05). Exposure to halothane is not recommended for use in female mice prior to mating procedures because it leads to decreased embryo production...(AU)

Embora os efeitos negativos dos anestésicos inalatórios na fertilidade já foram descritos há algum tempo, os estágios durante o ciclo reprodutivo em que estes efeitos ocorrem bem como os mecanismos de ação ainda permanecem desconhecidos. Os objetivos deste estudo foram avaliar os efeitos da exposição aguda em fêmeas de camundongos ao halotano, isofluorano, e sevofluorano prévio ao acasalamento. Fêmeas de camundongos BALB/c (n=160) foram alocadas em grupos de 20 aos tratamentos halotano (HG), isofluorano (IG), sevofluorano (SG), oxigênio (SH), e seus respectivos grupos controle (CGs). As fêmeas de camundongos foram expostas a uma concentração alveolar mínima (CAM) do anestésico correspondente ou a oxigênio por 4 horas diárias durante 5 dias consecutivos. Após dois dias do fim da exposição às fêmeas foram acasaladas com os machos (proporção 2:1 fêmea/macho) durante 5 dias consecutivos. A cada manhã, as fêmeas foram avaliadas para a observação da presença de plug vaginal. Metade das fêmeas que exibiam plug foram submetidas à eutanásia após 72 horas para avaliação embrionária. As fêmeas restantes foram eutanasiadas no 14º dia de gestação para avaliação fetal. No HG foi observado um menor número de mórulas e embriões totais (mórulas + blastocistos) (P 0,05). O número de implantações foi menor no HG (6,0) comparado ao IG (11,8) e SG (12,4) (P 0,05). A exposição ao halotano não...(AU)