Early maturation of corpus luteum in rabbits - effect of sildenafil citrate on luteolytic capacity in the early luteal period

Background: Prostaglandin F2 alpha (PGF2 α) binds to the specific receptor (PTGFR) on the corpus luteum (CL) in mammals, inducing regression of the CL structure (luteolysis) and initiating a new cycle. While PGF2 α is effective only on mature CL, the immature CL structure (early luteal phase) resists PGF2 α. In this study, sildenafil citrate, which is used to increase blood flow in the genital organs for treating specific pregnancy issues in women, was administered during the early luteal phase in a rabbit model to test the hypothesis of enhancing blood flow to the CL, thereby promoting earlier maturation and enabling a response to PGF2 α. Materials, Methods & Results: The study was conducted in 2 sub-studies: clinical and molecular. A large number of rabbits were initially included in the sub-studies to ensure a sufficient number of pseudo-pregnant rabbits. Ovulation in rabbits was induced with buserelin acetate and was considered as day 0 of the study. The sub-studies were continued with rabbits whose pseudo-pregnancies were confirmed according to progesterone (P4 ) results. As a result, the studies were continued with a total of 41 pseudo-pregnant New Zealand female rabbits, 21 of which were included in the clinical sub-study and 20 in the molecular sub-study. In both sub-studies, on day 3 of the luteal period, rabbits in the treatment group received 5 mg/kg sildenafil citrate and all rabbits received a single dose of exogenous PGF2 α on day 4 to induce luteolysis. In the clinical sub-study, echotexture and intraovarian blood flow changes in the ovaries were determined by ultrasonography (USG) examination. In the molecular sub-study, the expression changes of Hypoxia Inducible Factor 1 Alpha (HIF1A) and Vascular Endothelial Growth Factor (VEGF) related to angiogenesis, Steroidogenic Acute Regulatory Protein (StAR) related to P4 metabolism, Prostaglandin-Endoperoxide Synthase 2 (PTGS2) related to prostaglandin (PG) mechanism and 15-Hydroxyprostaglandin Dehydrogenase (HPGD) genes at mRNA level were determined using Real Time Polymerase Chain Reaction (RT-PCR) in CL tissues obtained with ovariohysterectomy (OVH) at 1 and 12 h after PGF2 α injection. In addition, blood samples were collected for determine P4 levels from all rabbits. In the clinical sub-study; there was no difference between the groups in mean gray values (MGV), whereas there was a significant decrease in both pulsatile index (PI) and resistance index (RI) values at 40 min after PGF2 α injection (P < 0.05). In the molecular sub-study, it was determined that sildenafil citrate had no significant effect (P > 0.05) on the expression levels 1 and 12 h after PGF2 α injection. According to the results of the molecular sub-study, no significant effect of sildenafil citrate on the mRNA expression levels in the investigated genes was detected (P > 0.05). However, within each group, differences were found according to OVH time after PGF2 α injection. It was observed that PTGS2 and HPGD mRNA expressions decreased at the 12th h compared to the 1st h, while HIF1A expression increased (P < 0.05). Discussion: According to the results obtained from clinical and molecular sub-studies, it was determined that a single dose of sildenafil citrate (5 mg/kg) applied on the 3rd day of the luteal period did not contribute to the maturation process of the CL, did not increase blood flow, and was insufficient to break the resistance of the CL against PGF2 α applied on the 4th day of the luteal period. However, a significant decrease in the PI value at the 40th min after PGF2 α injection suggests that sildenafil citrate has a supportive effect, and that this decrease is also seen in the RI value, suggesting that its effect is insufficient against the vasoconstrictive effect of PGF2 α.

Meta-analytical approaches to assess citric and ascorbic acid use in nursery piglets / Abordagens meta-analíticas para avaliar o uso do ácido cítrico e ascórbico em leitões em creche

This study evaluated different meta-analytical approaches tousing citric and ascorbic acids in the performance of nursery piglets. The study included 24 publications between 1985 and 2018 (mode 1998), totaling 4215 nursery piglets. The Higgins index indicated high heterogeneity (98.5%) among the studies concerning citric acid (96.9%) and ascorbic acid (91.4%). In variances and forest plot analysis, using citric and ascorbic acids and their analogs was no significant effect (P > 0.05) on piglet weight gain. In the meta-analysis approach proposed by LOVATTO et al. (2007) and SAUVANT et al. (2008), there was no significant effect (P > 0.05) of the addition of citric and ascorbic acids on piglet performance. Numerically, was observed an increased ADG (Δ > 0) related to acids (supplemented over control treatments) in 59.7% (ascorbic acid) and 46.6% (citric acid) of the ADG comparisons. The effect of ascorbic supplementation on the respective control treatment of the overall nursery period was on average 0.98%, 1.07%, and -1.3% for ADFI, ADG, and FCR, respectively. Citric acid supplementation to the respective control treatment of the overall nursery period was on average 1.67%, 4.40%, and -2.29% for ADFI, ADG, and FCR. Adding citric and ascorbic acids in diets for nursery piglets does not alter performance regardless of the meta-analysis method used.

Neste estudo, o objetivo foi avaliar diferentes abordagens de sistematização e meta-análise sobre o uso de ácidos cítrico e ascórbico no desempenho de leitões de creche. O estudo incluiu 24 publicações entre 1985 e 2018 (moda 1998), totalizando 4215 leitões de creche. O índice de Higgins indicou alta heterogeneidade entre os estudos envolvendo ácido cítrico (96,9%) e ácido ascórbico (91,4%). Na análise por forest plot a adição de ácido ascórbico em dietas de leitões não melhora (P > 0.05) o ganho de peso de leitões em creche. Na abordagem de meta-análise proposta por LOVATTO et al. (2007) e SAUVANT et al. (2008) não houve efeito significativo (P > 0.05) da adição de ácidos cítrico e de ascórbico no desempenho dos leitões. Numericamente, observou-se aumento do ganho de peso (Δ > 0) relacionado aos ácidos (suplementado em relação aos tratamentos controle) observado em 59.7% (acido ascórbico) e 46.6% (ácido cítrico) das comparações do ganho de peso diário. A suplementação de ácido ascórbico em relação ao tratamento controle no período total de creche teve variação de 0,98%, 1,07% e -1,3% para ADFI, ADG e FCR, respectivamente. A adição de ácido cítrico em relação ao tratamento controle teve variação de 1,67%, 4,40% e -2,29 para ADFI, ADG e FCR. A adição de ácidos cítrico e ascórbico em dietas para leitões de creche não altera o desempenho, independentemente da abordagem de meta-análise utilizada.

Canine sperm vitrification with nonpermeable cryoprotectants and coconut water extender

This study was aimed to assess the efficiency of coconut water extender with addition of soy lecithin and sucrose as nonpermeable cryoprotectants for canine semen vitrification, using a simple method that yields a high survival rate of spermatozoa for clinical use. Twelve ejaculates from 12 adult normozoospermic dogs were collected separately by digital manipulation and only the second semen fraction was used in this study. After evaluation of volume, concentration, viability, total and progressive motility, velocity parameters and morphology, semen was diluted with a coconut water extender (50% (v/v(volume per volume)) coconut water, 25% (v/v) distilled water and 25% (v/v) 5% anhydrous monosodium citrate solution) with addition of soy lecithin and fructose at 1% and 0.25M sucrose until final concentration of 100x106 spermatozoa/ml. After equilibration at 5ºC for 60 minutes, semen was vitrified by "direct dropping method" into liquid nitrogen in spheres with a volume of 30 µl. After a week of storage the spheres were devitrified as three of them were dropped into 0.5 mL of CaniPlus AI medium (Minitüb, Germany), which was previously warmed in a water bath at 42ºC for 2 minutes and evaluated about the above mentioned parameters. It was found that vitrification resulted in a lower percentage of viable sperms, normal morphology, total and progressive motilities (p0.05) compared to fresh semen samples. In conclusion, our results demonstrate that vitrification with coconut water extender with addition of 1% soy lecithin and 0.25M sucrose as cryoprotectants, has an excellent potential for routine canine sperm cryopreservation.(AU)

Use of phytase and citric acid supplementation on growth performance and nutrient digestibility of Cirrhinus mrigala fingerlings fed on canola meal based diet / Uso de suplementação de fitase e ácido cítrico no desempenho do crescimento e digestibilidade de nutrientes de dedos Cirrhinus mrigala alimentados em dieta à base de refeição de canola

Fishmeal; being a limited and costly feed ingredient is continuously been substituted with locally available plant proteins. However, the occurrence of anti-nutritional factors in plant meal suppresses its potential to be fully replaced. Therefore, in this study we aimed to study the synergistic effects of dietary additives like citric acid and phytase enzyme supplementation on growth performance and nutrient digestibility of Cirrhinus mrigala fingerlings. Canola meal (CM) was used as a test ingredient to replace fishmeal (FM) as; 0%, 25%, 50% and 75%. These four diets were further supplemented by varying levels of phytase (0 and 750 FTU kg-1) and citric acid (0% and 2.5%) to formulate total sixteen test diets as T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15 and T16. Each treatment contained three replicates; applied to fish groups having 15 fingerlings each; following 3×3 factorial arrangement. 1% of chromic oxide was added as an inert marker. Maximum weight gain% (288%) and the lowest value of FCR (1.07) were recorded when fish was fed on diet T12 as compared to fish fed control diet (T1). Similarly, optimum nutrient digestibility values such as crude protein (77%), crude fat (84%) and gross energy (70%) were noted on same level. It was concluded that 50% canola meal can optimally replace fishmeal when supplemented with phytase and citric acid at the levels of 750 FTU kg-¹ and 2.5%, respectively.

A farinha de peixe, por ser um ingrediente alimentar limitado e caro, é continuamente substituída por proteínas vegetais disponíveis localmente. No entanto, a ocorrência de fatores antinutricionais na farinha de plantas suprime seu potencial de ser totalmente substituída. Portanto, neste estudo objetivamos estudar os efeitos sinérgicos de aditivos dietéticos como ácido cítrico e suplementação com enzima fitase sobre o desempenho de crescimento e digestibilidade de nutrientes de alevinos de Cirrhinus mrigala. A farinha de canola (CM) foi usada como ingrediente de teste para substituir a farinha de peixe (FM) como: 0%, 25%, 50% e 75%. Essas quatro dietas foram suplementadas por níveis variados de fitase (0 e 750 FTU kg-1) e ácido cítrico (0% e 2,5%) para formular um total de 16 dietas de teste como T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15 e T16. Cada tratamento continha três repetições; aplicado a grupos de peixes com 15 alevinos cada; seguindo o arranjo fatorial 3 × 3. 1% de óxido crômico foi adicionado como um marcador inerte. % de ganho de peso máximo (288%) e o valor mais baixo de FCR (1,07) foram registrados quando os peixes foram alimentados com dieta T12 em comparação com peixes alimentados com dieta controle (T1). Da mesma forma, valores ótimos de digestibilidade de nutrientes, como proteína bruta (77%), gordura bruta (84%) e energia bruta (70%) foram anotados no mesmo nível. Concluiu-se que 50% da farinha de canola pode substituir de forma ideal a farinha de peixe quando suplementada com fitase e ácido cítrico nos níveis de 750 FTU kg-¹ e 2,5%, respectivamente.

Use of phytase and citric acid supplementation on growth performance and nutrient digestibility of Cirrhinus mrigala fingerlings fed on canola meal based diet / Uso de suplementação de fitase e ácido cítrico no desempenho do crescimento e digestibilidade de nutrientes de dedos Cirrhinus mrigala alimentados em dieta à base de refeição de canola

Abstract Fishmeal; being a limited and costly feed ingredient is continuously been substituted with locally available plant proteins. However, the occurrence of anti-nutritional factors in plant meal suppresses its potential to be fully replaced. Therefore, in this study we aimed to study the synergistic effects of dietary additives like citric acid and phytase enzyme supplementation on growth performance and nutrient digestibility of Cirrhinus mrigala fingerlings. Canola meal (CM) was used as a test ingredient to replace fishmeal (FM) as; 0%, 25%, 50% and 75%. These four diets were further supplemented by varying levels of phytase (0 and 750 FTU kg-1) and citric acid (0% and 2.5%) to formulate total sixteen test diets as T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15 and T16. Each treatment contained three replicates; applied to fish groups having 15 fingerlings each; following 3×3 factorial arrangement. 1% of chromic oxide was added as an inert marker. Maximum weight gain% (288%) and the lowest value of FCR (1.07) were recorded when fish was fed on diet T12 as compared to fish fed control diet (T1). Similarly, optimum nutrient digestibility values such as crude protein (77%), crude fat (84%) and gross energy (70%) were noted on same level. It was concluded that 50% canola meal can optimally replace fishmeal when supplemented with phytase and citric acid at the levels of 750 FTU kg-1 and 2.5%, respectively.

Resumo A farinha de peixe, por ser um ingrediente alimentar limitado e caro, é continuamente substituída por proteínas vegetais disponíveis localmente. No entanto, a ocorrência de fatores antinutricionais na farinha de plantas suprime seu potencial de ser totalmente substituída. Portanto, neste estudo objetivamos estudar os efeitos sinérgicos de aditivos dietéticos como ácido cítrico e suplementação com enzima fitase sobre o desempenho de crescimento e digestibilidade de nutrientes de alevinos de Cirrhinus mrigala. A farinha de canola (CM) foi usada como ingrediente de teste para substituir a farinha de peixe (FM) como: 0%, 25%, 50% e 75%. Essas quatro dietas foram suplementadas por níveis variados de fitase (0 e 750 FTU kg-1) e ácido cítrico (0% e 2,5%) para formular um total de 16 dietas de teste como T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15 e T16. Cada tratamento continha três repetições; aplicado a grupos de peixes com 15 alevinos cada; seguindo o arranjo fatorial 3 × 3. 1% de óxido crômico foi adicionado como um marcador inerte. % de ganho de peso máximo (288%) e o valor mais baixo de FCR (1,07) foram registrados quando os peixes foram alimentados com dieta T12 em comparação com peixes alimentados com dieta controle (T1). Da mesma forma, valores ótimos de digestibilidade de nutrientes, como proteína bruta (77%), gordura bruta (84%) e energia bruta (70%) foram anotados no mesmo nível. Concluiu-se que 50% da farinha de canola pode substituir de forma ideal a farinha de peixe quando suplementada com fitase e ácido cítrico nos níveis de 750 FTU kg-1 e 2,5%, respectivamente.

Use of phytase and citric acid supplementation on growth performance and nutrient digestibility of Cirrhinus mrigala fingerlings fed on canola meal based diet / Uso de suplementação de fitase e ácido cítrico no desempenho do crescimento e digestibilidade de nutrientes de dedos Cirrhinus mrigala alimentados em dieta à base de refeição de canola

Fishmeal; being a limited and costly feed ingredient is continuously been substituted with locally available plant proteins. However, the occurrence of anti-nutritional factors in plant meal suppresses its potential to be fully replaced. Therefore, in this study we aimed to study the synergistic effects of dietary additives like citric acid and phytase enzyme supplementation on growth performance and nutrient digestibility of Cirrhinus mrigala fingerlings. Canola meal (CM) was used as a test ingredient to replace fishmeal (FM) as; 0%, 25%, 50% and 75%. These four diets were further supplemented by varying levels of phytase (0 and 750 FTU kg-1) and citric acid (0% and 2.5%) to formulate total sixteen test diets as T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15 and T16. Each treatment contained three replicates; applied to fish groups having 15 fingerlings each; following 3×3 factorial arrangement. 1% of chromic oxide was added as an inert marker. Maximum weight gain% (288%) and the lowest value of FCR (1.07) were recorded when fish was fed on diet T12 as compared to fish fed control diet (T1). Similarly, optimum nutrient digestibility values such as crude protein (77%), crude fat (84%) and gross energy (70%) were noted on same level. It was concluded that 50% canola meal can optimally replace fishmeal when supplemented with phytase and citric acid at the levels of 750 FTU kg-¹ and 2.5%, respectively.(AU)

A farinha de peixe, por ser um ingrediente alimentar limitado e caro, é continuamente substituída por proteínas vegetais disponíveis localmente. No entanto, a ocorrência de fatores antinutricionais na farinha de plantas suprime seu potencial de ser totalmente substituída. Portanto, neste estudo objetivamos estudar os efeitos sinérgicos de aditivos dietéticos como ácido cítrico e suplementação com enzima fitase sobre o desempenho de crescimento e digestibilidade de nutrientes de alevinos de Cirrhinus mrigala. A farinha de canola (CM) foi usada como ingrediente de teste para substituir a farinha de peixe (FM) como: 0%, 25%, 50% e 75%. Essas quatro dietas foram suplementadas por níveis variados de fitase (0 e 750 FTU kg-1) e ácido cítrico (0% e 2,5%) para formular um total de 16 dietas de teste como T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15 e T16. Cada tratamento continha três repetições; aplicado a grupos de peixes com 15 alevinos cada; seguindo o arranjo fatorial 3 × 3. 1% de óxido crômico foi adicionado como um marcador inerte. % de ganho de peso máximo (288%) e o valor mais baixo de FCR (1,07) foram registrados quando os peixes foram alimentados com dieta T12 em comparação com peixes alimentados com dieta controle (T1). Da mesma forma, valores ótimos de digestibilidade de nutrientes, como proteína bruta (77%), gordura bruta (84%) e energia bruta (70%) foram anotados no mesmo nível. Concluiu-se que 50% da farinha de canola pode substituir de forma ideal a farinha de peixe quando suplementada com fitase e ácido cítrico nos níveis de 750 FTU kg-¹ e 2,5%, respectivamente.(AU)

Curcumin alleviated oxidation stress injury by mediating osteopontin in nephrolithiasis rats

Purpose: To explore the role and mechanism of curcumin (Cur) in reducing oxidative stress damage in rats with nephrolithiasis induced by ethylene glycol (EG). Methods: Thirty male rats were divided into normal control, model, positive (10% potassium citrate), Cur-10 (10 mg/kg curcumin) and Cur-20 (20 mg/kg curcumin) groups. Results: The results of kidney tissue section stained by hematoxylin-eosin and von Kossa showed that curcumin treatment can inhibit the formation of kidney stones. The biochemical test results showed that the urea (Ur), creatinine (Cr), uric acid (UA), inorganic phosphorus and Ca2+ concentrations in urine decreased after being treated with curcumin. There were significant differences between different doses of curcumin (P < 0.05). Compared with the Cur-10 group, Cur-20 had a more significant inhibitory effect on malondialdehyde (MDA) (P < 0.05). In addition, reverse transcription polymerase chain reaction (PCR) detection and immunohistochemical results indicated that the osteopontin (OPN) in the kidney was significantly reduced after curcumin treatment. Conclusion: Curcumin could reduce the oxidative stress damage caused by EG-induced kidney stones.

Comparative study between S-(+)-ketamine­midazolam and fentanyl­droperidol in black-tufted marmosets (Callithrix penicillata) / Estudo comparativo entre S-(+)cetamina-midazolam e fentanil-droperidol em saguis-de-tufo-preto (Callithrix penicillata)

Sedative and antinociceptive effects of two anesthetic protocols in black-tufted marmosets were compared in this study. Twenty-six marmosets underwent chemical immobilization for physical examination, blood sampling, tattooing, and microchipping. Animals were randomly treated with S-(+)-ketamine (10 mg/kg) and midazolam (1 mg/kg) (KM) or fentanyl (12.5 µg/kg) and droperidol (625 µg/kg) (FD) given by intramuscular injection. Heart and respiratory rates were recorded. Sedation, antinociception, muscle relaxation, posture, auditory, and visual responses were evaluated using a scoring system. Sedation in KM was achieved faster (p < 0.001) and lasted for a shorter period of time (p = 0.0009). KM was similar to FD in its cardiorespiratory effects, auditory and visual responses. Both protocols promoted adequate sedation to allow manipulation. Animals in KM assumed lateral recumbency while animals in FD maintained a quadrupedal posture during evaluation. FD produced less intense sedation and muscle relaxation but a higher degree of antinociception compared to KM and is suitable for procedures that require analgesia in black-tufted marmosets.(AU)

O presente estudo comparou os efeitos cardiorrespiratórios, sedativos e antinociceptivos de dois protocolos anestésicos em saguis-de-tufo-preto (Callithrix penicillata). Vinte e seis saguis foram submetidos à contenção química para exame físico, coleta de sangue, tatuagem de identificação e microchip. Os animais foram tratados aleatoriamente com a associação de S-(+)-cetamina (10 mg/kg) e midazolam (1 mg/kg) (KM) ou fentanil (12,5 µg/kg) e droperidol (625 µg/kg) (FD), administrados por injeção intramuscular. Foram avaliadas frequência cardíaca, frequência respiratória, sedação, antinocicepção, relaxamento muscular, postura e resposta ao estímulo auditivo e visual. A sedação em KM foi alcançada mais rapidamente (p <0,001) e teve um tempo hábil mais curto (p = 0,0009). KM foi semelhante a FD nos efeitos cardiorrespiratórios, respostas auditivas e visuais. Os dois protocolos promoveram sedação adequada para manipulação. Os animais do grupo KM permaneceram em decúbito lateral durante a avaliação, enquanto os animais em FD mantiveram postura quadrupedal. FD resultou em sedação e relaxamento muscular de menor intensidade, porém com maior escore de antinocicepção em comparação com KM, sendo adequada para procedimentos que requerem analgesia em saguis-de-tufo-preto.(AU)

Anticoagulants and their effects on the hematological and biochemical parameters of yellow-spotted amazon river turtle / Anticoagulantes e seus efeitos em parâmetros hematológicos e bioquímicos de tracajá

Knowledge of suitable methods and reagents for assessing the health condition of specimens of a given species is essential. The present study evaluated the efficacy of the heparin anticoagulants 5,000 I.U. mL-1, Na2EDTA, and K3EDTA on the blood parameters of yellow-spotted amazon river turtle Podocnemis unifilis, employing different solutions for red blood cells count. The use of the various anticoagulants evaluated after 10 hours of storage efficiently inhibited coagulation in blood samples from P. unifilis. An increased number of erythrocytes was observed with the use of K3EDTA 5% when compared with heparin. Statistically significant changes in the erythrocyte number were observed with the use of the different solutions. Solutions which featured sodium citrate and formaldehyde in their composition, allowed erythrocytes counting up to 120 hours after blood collection, without a change in values. The use of the heparin anticoagulants 5,000 I.U. mL-1, Na2EDTA 3%, Na2EDTA 5%, K3EDTA 3% was recommended in the hematological analysis of P. unifilis. Also recommended was the use of the formaldehyde-citrate solution containing 1.9 g of sodium citrate and 1.0 mL of formaldehyde (in 50 mL of distilled water) to perform red blood cells counts in yellow-spotted amazon river turtle.

O conhecimento sobre métodos e reagentes apropriados para as avaliações da condição de saúde de exemplares de determinada espécie é fundamental. Este estudo avaliou a eficácia dos anticoagulantes heparina 5.000 U.I. mL-1, Na2EDTA (3% e 5%), e K3EDTA (3 e 5%) em parâmetros sanguíneos de tracajá (Podocnemis unifilis), bem como diferentes soluções para contagem de eritrócitos totais. A coagulação foi eficientemente inibida nas amostras de sangue de P. unifilis com o uso dos diferentes anticoagulantes avaliados após 10 horas de armazenamento. Maior número de eritrócitos com o uso de K3EDTA 5% foi observado quando comparado com a coleta de sangue realizada com heparina. Diferenças, estatisticamente significativas, entre as contagens de eritrócitos com o uso das diferentes soluções de reagentes avaliadas foram verificadas. As soluções contendo citrato de sódio e formol na composição possibilitaram contagens de eritrócitos até 120 horas após a coleta, sem alteração em seus valores. Recomenda-se o uso dos anticoagulantes heparina 5000 U.I. mL-¹, Na2EDTA 3%, Na2EDTA 5%, K3EDTA 3% nas análises hematológicas de tracajá. Assim como o uso da solução de formol-citrato contendo 1,9 g de citrato de sódio e 1,0 mL de formol (em 50 mL de água destilada) para realização das contagens de eritrócitos totais em tracajá.

Comparative effectiveness of EDTA and citric acid assisted phytoremediation of Ni contaminated soil by using canola (Brassica napus) / Eficácia comparativa da fitorremediação assistida por EDTA e ácido cítrico de solo contaminado com Ni, usando canola (Brassica napus)

Phytoremediation is an ecofriendly technique to clean heavy metals from contaminated soil by the use of high biomass producing plant species. Chelators can help to improve this biological technique by increasing metal solubility. Therefore, a pot experiment was conducted to determine the effect of the chelators EDTA and citric acid (CA) in phytoremediation of Ni contaminated soil by using Brassica napus (canola). Two cultivars of B. napus, Con-II (tolerant) and Oscar (sensitive), were selected after screening and exposed to NiSO4 at 30 ppm at the time of sowing. CA (10 mM) and EDTA (1.5 mM) were applied either alone or in combination with each other after two weeks of Ni treatments. Different parameters like morpho-physiological and biochemical data were recorded after 15 days of chelate application. The results highlighted the successful use of chelating agents (CA and EDTA) not only to ameliorate Ni stress but also to enhance Ni accumulation which is prerequisite for phytoremediation. The basal application of 10 mMCA and 1.5 mM EDTA concentration proved to be effective for the growth of plants. The combination of chelating agents failed to show any synergistic effects.(AU)

A fitorremediação é uma técnica ecologicamente correta para limpar metais pesados ​​de solo contaminado pelo uso de espécies vegetais produtoras de alta biomassa. Os quelantes podem ajudar a melhorar esta técnica biológica aumentando a solubilidade do metal. Para tanto, foi realizado um experimento em vaso para determinar o efeito dos quelantes EDTA e ácido cítrico (AC) na fitorremediação de solo contaminado com Ni, utilizando Brassica napus (canola). Duas cultivares de B. napus, Con-II (tolerante) e Oscar (sensível) foram selecionadas após triagem e expostas a NiSO4 a 30 ppm no momento da semeadura. CA (10 mM) e EDTA (1,5 mM) foram aplicados sozinhos ou em combinação um com o outro após duas semanas de tratamentos com Ni. Diferentes parâmetros como dados morfofisiológicos e bioquímicos foram registrados após 15 dias de aplicação de quelato. Os resultados destacaram o uso bem-sucedido de agentes quelantes (CA e EDTA) não apenas para melhorar o estresse de Ni, mas também para aumentar o acúmulo de Ni, um pré-requisito para a fitorremediação. A aplicação basal da concentração de 10 mMCA e 1,5 mM de EDTA mostrou-se eficaz para o crescimento das plantas. A combinação de agentes quelantes não mostrou quaisquer efeitos sinérgicos.(AU)

The Effects of Saccharomyces Cerevisiae and Citric Acid on Productive Performance, Egg Quality Parameters, Small Intestinal Morphology, and Immune-Related Gene Expression in Laying Japanese Quails

This experiment evaluated the effects of Saccharomyces cerevisiae (S. cerevisiae) and citric acid on production performance, egg quality, intestine histomorphology, and avian ß-defensin 1 and 2 (AvBD 1 and 2) gene expressions in laying Japanese quails. A total of 400 48-day-old quails were randomly assigned to a 2×2×2 factorial arrangement of treatments with 5 replicates (each containing 10 quails) for 7 weeks. Variable factors consisted of S. cerevisiae (0 and 100 mg/kg diet), citric acid (0 and 5 g/kg diet), and Virginiamycin (0 and 50 mg/kg diet). At the completion of the trial, one bird per replicate was randomly killed, and jejunal tissue samples were removed to evaluate intestinal morphometric characteristics. Samples were taken from the midpoint of the jejunum to measure the gene expression of AvBD 1 and 2. Dietary inclusion of both S. cerevisiae and citric acid resulted in increased egg weight, egg mass, reduced feed intake, and improved FCR (p<0.05). The addition of S. cerevisiae to diets containing citric acid reduced feed intake, increased egg weight, and improved FCR (p<0.05). Shell weight and shell thickness were increased in birds fed each of S. cerevisiae and citric acid supplements (p<0.05). Dietary S. cerevisiae and citric acid similarly increased intestinal villus height, width, surface area, and the villus height to crypt depth ratio (p<0.0001). Results showed that AvBD 1 and 2 genes expression were up-regulated on quails fed S. cerevisiae-supplemented diets (p<0.0001). In conclusion, these results suggest that supplementation of S. cerevisiae and citric acid as functional feed additives either alone or in combination could be a potential alternative to antibiotics in the diet of Japanese laying quails.(AU)

Canine cystitis - biofilm formation by bacterial isolates

Background: Biofilms have been reported as important virulent markers associated with drug resistance in urinary tractinfections (UTIs) in humans and dogs. However, in veterinary medicine, researches involving biofilm formation, treatments and preventions have been limited; yet, it is still possible to find few studies demonstrating biofilm-forming bacteriaassociated with different comorbidities such as otitis, wound infections, UTIs, and endometritis. These studies generallyselect dogs with chronic and recurrent infections, which could be an important factor in antibiotic resistance. We aimed toevaluate biofilms in sporadic cystitis regarding prevalence and drug resistance.Materials, Methods & Results: Urine samples were collected by cystocentesis from 36 client-owned dogs under clinicaland laboratory suspicion of non-recurrent urinary bladder infection (cystitis). Urine was aseptically plated onto bloodagar, MacConkey, and CLED, followed by incubation for 24 to 48 h. Definitive identification of a potential pathogen wasmade by subculture collected from an isolated colony to obtain a pure culture. The gram staining method and specificbiochemical tests (phenol red fermentation, lysine, phenylalanine, citrate, sulfide-indole-motility, and urease) were usedto distinguish and classify the bacteria. After identification, the bacteria were tested for antimicrobial susceptibility by astandard disk diffusion method, using the following antimicrobials: amoxicillin with clavulanic acid, ampicillin, ceftriaxone, ciprofloxacin, clindamycin, cefazolin, cephalothin, erythromycin, gentamicin, norfloxacin, and sulfamethoxazoletrimethoprim. The biofilm-forming ability was determined based on a culture...

Infusão contínua de fentanil e cetamina para controle de dor no transcirurgico: relato de caso / Continuous infusion of fentanyl and ketamine for trans-surgical pain control: case report

O paciente da espécie canina, sem raça definida, com 5 anos, deu entrada a clínica veterinária particular apresentando quadro de dor aguda e intensa em cavidade abdominal associada a prostração. Foram realizados exames de sangue, ultrassonografia e radiografia de abdômen, cuja suspeita foi torção parcial do estômago, com esplenomegalia, além de gastrite e enterite. O paciente foi submetido imediatamente ao procedimento cirúrgico para alívio, reposicionamento do órgão e gastropexia. O animal foi medicado previamente com metadona, tramadol e dipirona duas horas antes do ato cirúrgico, portanto não foram utilizadas medicações pré-anestésicas e optou-se pelo propofol e cetamina para a indução anestésica, além da infusão contínua com cetamina e fentanil para se obter a analgesia multimodal. Os parâmetros de dor foram avaliados por meio das frequências cardíaca e respiratória, traçado eletrocardiográfico, pressão arterial sistólica e média, aferição da temperatura retal e coloração de mucosas, a cada 5 minutos. Os mesmos indicaram boa estabilidade hemodinâmica, segurança e analgesia, sem necessidade de alterar as doses estabelecidas durante o procedimento. Pode ser observada também redução do requerimento de isoflurano para manutenção do plano cirúrgico. O paciente acordou sem sinais de dor no pós-operatório imediato evitando estresse sistêmico, o que conferiu o bom resultado do protocolo analgésico instituído.

The patient (Caramel) canine, mixed breed, approximately 5 years old, weighing 26kg was seen at the Ame Vet veterinary clinic, with severe abdominal pain, apathy, anorexia, prostration. Blood tests, abdominal ultrasound and abdominal x-ray were performed according to the tests a suspicion of partial torsion of the stomach and splenomegaly with right displacement, in addition to gastritis and enteritis. It was decided by the clinic and surgeon to perform laparotomy and prophylactic gastropexy. The animal was previously medicated by the clinic that treated the animal for analgesia with methadone, tramadol and dipyrone two hours before the beginning of the surgical procedure, therefore, pre-anesthetic medications, anesthetic induction with propofol, and ketamine were not used, aiming to perform analgesia. Multimodal with continuous infusion of ketamine and fentanyl, that is, more than one different mechanism of action, acting synergistically and reducing dose-dependent adverse effects and monitoring hemostasis and patient parameters, presenting its efficacy and safety in intraoperative pain control. The parameters addressed were heart rate (HR), respiratory rate (RR), electrocardiographic tracing, Spo2, systolic blood pressure (SBP), mean arterial pressure (MAP), temperature and mucosal color, evaluated every 5 minutes. The parameters evaluated indicated good hemodynamic stability, safety and analgesia, with no need to change the doses established during the procedure. A reduction in the requirement of isoflurane to maintain the surgical plan can also be observed. The patient woke up without signs of pain in the immediate postoperative period, thus avoiding systemic stress. which conferred the good result of the analgesic protocol instituted.

Analgesia pós-operatória com Fentanil transdérmico ou Metadona intramuscular em cães submetidos à hemilaminectomia toracolombar / Postoperative analgesia with transdermal Fentanyl or intramuscular Methadone in dogs submitted to thoracolumbar hemilaminectomy

Background: Postoperative care after thoracolumbar hemilaminectomy in dogs generally includes rest, physical therapy,and analgesics such as opioids. Currently, there is no established standard for the management of postoperative pain inpatients undergoing hemilaminectomy. Ideally, an analgesic protocol should provide adequate pain relief with limited sedation, low adverse effects, and postoperative patient comfort. Therefore, this study aimed to evaluate postoperative analgesiausing transdermal fentanyl or intramuscular methadone in dogs undergoing thoracolumbar hemilaminectomy associatedwith intervertebral disc fenestration (HT) for the treatment of intervertebral disc extrusion (IVDE).Materials, Methods & Results: Eight dogs from the Department of Neurology and Neurosurgery of a Veterinary TeachingHospital, submitted to HT for the treatment of IVDE, were included. The dogs were randomly distributed into 2 groups ofequal numbers, namely the transdermal fentanyl (FT) group and the intramuscular methadone (IM) group. At the end ofthe surgical procedure, a fentanyl adhesive patch was applied to the animals in the FT group, which remained there for 72h. In the IM group, analgesia was induced by intramuscular administration of methadone at intervals of 6 h until 72 h aftersurgery. The animals were evaluated using the short form of the Glasgow Composite Measure Pains Scale (CMPS-SF).Evaluations of physiological parameters, side effects, and pain were performed by 2 assessors who had experience usingthe pain scale and were blinded to the analgesic protocol. Pain evaluations were performed every 2 h (from T4) until 24h after the surgical procedure. Evaluations were performed every 4 h from 24 h to 48 h after the surgical procedure and...

Evaluating chemicals for inducing budburst in peach orchard grown in subtropical conditions / Avaliação de produtos químicos para indução de brotação em pomar de pessegueiro cultivado em condições subtropicais

The availability of chill hours influences the break dormancy, sprouting and production of temperate fruit trees in different regions. During the winter, the chilling requirements for growing peaches in subtropical locations are restricted, and certain chemicals are required to overcome dormancy and induce budburst. As a result, further information concerning the use of these compounds on stimulating sprouting in peach trees is required. The purpose of this study was to evaluate the ability of hydrogen cyanamide, nitrogen fertilizer, and calcium nitrate to trigger budburst in the 'Douradão' peach orchard, which is in the subtropical area of São Paulo state, Brazil. The following tree chemicals were tested: 0.6 % hydrogen cyanamide, 2.5 % nitrogen fertilizer with 4 % calcium nitrate, 4 % calcium nitrate, and control. The hydrogen cyanamide accelerated, predicted, and focused blooming on plants, along with improving fruit set and output. Furthermore, combining nitrogen fertilizer with calcium nitrate accelerated budburst, which boosted flowering, output per tree, and yield. The spraying of the peach plants with just calcium nitrate produced results like those obtained in the control. The results allow for the use of hydrogen cyanamide or nitrogen fertilizer + calcium nitrate as an option for triggering budburst in Douradão cultivar peach trees under subtropical circumstances, depending on the orchards system management used.(AU)

A disponibilidade de horas de frio influencia a quebra da dormência, brotação e produção de árvores frutíferas de clima temperado em diferentes regiões. Os requerimentos de frio para o cultivo de pessegueiros em regiões subtropicais são limitados durante o inverno e alguns produtos químicos são necessários para superar a dormência e induzir a brotação. Portanto, são necessárias mais informações sobre o uso desses compostos na promoção da brotação em pessegueiros. Este trabalho teve como objetivo avaliar a cianamida hidrogenada, o fertilizante nitrogenado e o nitrato de cálcio para brotação, num pomar de pessegueiro 'Douradão', localizado em região subtropical do estado de São Paulo, Brasil. Para isso, três produtos químicos foram avaliados: 0,6% de cianamida hidrogenada; 2,5% de fertilizante nitrogenado + 4% de nitrato de cálcio; 4% de nitrato de cálcio e o controle. A cianamida hidrogenada aumentou o florescimento das plantas, além de aumentar a frutificação e a produção. Além disso, fertilizante nitrogenado + nitrato de cálcio também proveram a brotação, potencializando o florescimento e a produção das plantas. A pulverização dos pessegueiros somente com nitrato de cálcio apresentou desempenho semelhante ao observado na testemunha. Os resultados permitiram indicar a utilização da cianamida hidrogenada ou fertilizante nitrogenado + nitrato de cálcio, como alternativa para indução de brotação em pessegueiros cultivar Douradão, em condições subtropicais, dependendo do sistema de cultivo adotado nos pomares.(AU)

Molecular analysis of Bartonella spp. in liver tissue of bats from the Atlantic Forest biome, Brazil / Análise molecular de Bartonella spp. em tecido hepático de morcegos do bioma Mata Atlântica, Brasil

The genus Bartonella comprises gram-negative bacilli that possess tropism for erythrocytes and endothelial cells in animals of the orders Rodentia, Lagomorpha, Carnivora, Artiodactyla, Eulipotyphla, and Chiroptera. Bacterial infection may be associated with lymphadenitis, endocarditis, bacillary angiomatosis, and peliosis hepatis. Thirteen species of Bartonella are recognized as zoonotic and bats are considered to be their potential reservoirs. This study aimed to analyze the occurrence of Bartonella spp. in livers of neotropical bats belonging to the families Molossidae, Phyllostomidae, and Vespertilionidae, classified into 21 genera from the Atlantic Forest biome in São Paulo. A total of 341 (n = 341) chiropterans samples were tested for the presence of citrate synthase (gltA) gene of Bartonella by partial amplification using polymerase chain reaction (PCR). Samples of two bats (0.6%) of the species Glossophaga soricina from the municipality of São Roque were tested positive for gltA gene. Phylogenetic analysis indicated that sequences clustered in a clade that was close to the Bartonella sp. detected in G. soricina, which was collected from the Cerrado biome in Tocantins, Brazil. Despite a low prevalence of the detected infection, results indicated that neotropical bats from the Atlantic Forest were potential hosts of Bartonella spp., which might be related to the maintenance of a wild enzootic cycle of the bacterium. Additional studies, particularly on bats of genus Glossophaga, are required to elucidate the dynamics of intraspecific relationships between etiologic agent-vector-hosts.

O gênero Bartonella compreende bacilos gram-negativos que possuem tropismo por eritrócitos e células endoteliais em animais das ordens Rodentia, Lagomorpha, Carnivora, Artiodactyla, Eulipotyphla e Chiroptera. A infecção bacteriana pode estar associada a linfadenite, endocardite, angiomatose bacilar e peliose hepática. Treze espécies de Bartonella são reconhecidas como zoonóticas e os morcegos são considerados seus potenciais reservatórios. Este estudo teve como objetivo analisar a ocorrência de Bartonella spp. em fígados de morcegos neotropicais pertencentes às famílias Molossidae, Phyllostomidae e Vespertilionidae, classificados em 21 gêneros do bioma Mata Atlântica em São Paulo. Um total de 341 (n = 341) amostras de quirópteros foram testados para a presença do gene da citrato sintase (gltA) de Bartonella por amplificação parcial usando reação em cadeia da polimerase (PCR). Amostras de dois morcegos (0,6%) da espécie Glossophaga soricina do município de São Roque foram testadas positivas para o gene gltA. A análise filogenética indicou que as sequências agrupadas em um clado próximo ao de Bartonella sp. detectada em G. soricina, que foi coletada no bioma Cerrado no Tocantins, Brasil. Apesar da baixa prevalência da infecção detectada, os resultados indicaram que os morcegos neotropicais da Mata Atlântica eram potenciais hospedeiros de Bartonella spp., o que pode estar relacionado à manutenção de um ciclo enzoótico selvagem da bactéria. Estudos adicionais, particularmente em morcegos do gênero Glossophaga, são necessários para elucidar a dinâmica das relações intraespecíficas entre agente etiológico-vetor-hospedeiro.

Short chain organic acids in poultry: Literature review

The importance of Brazil for the International Trade of poultry products induces legislators and interested companies to pay special attention to complying with international standard quality requirements. One of these requirements is the suppression of the indiscriminate use of antibiotics in poultry, as in the strategy of their use as a growth promoter. To meet this demand, the study of short-chain organic acids becomes interesting and their review essential. As good examples for the issue described, short chain organic acids such as citric acid, fumaric acid, sorbic acid and malic acid are extremely important due to their antimicrobial qualities, accessibility, and international acceptance. In addition to the individual effect, the blend of acids, that is, the association of acids for the role of growth promoter or performance enhancer, combines acids that act synergistically, enhancing their qualities. This study aims to address the physicochemical characteristics, effects, and qualities of short-chain organic acids, as well as their associated effects, encouraging further work that fills the scientific gaps in empirical knowledge about these additives.(AU)

Systolic blood pressure in anesthetized bitches measured with photoplethysmography

Background: Photoplethysmography is widely used in human medicine, with few studies on its use in veterinary medicine. Its sensor detects fluctuations in blood volume at the site, providing direct readings of cardiac pulse and peripheral oxygen saturation, as well as estimating cardiac output, respiratory rate and blood pressure. This study aimed to evaluate the use of photoplethysmography and compare it to vascular Doppler ultrasound as an indirect method of measuring systolic blood pressure in bitches undergoing elective ovariohysterectomy, using the invasive assessment of systolic blood pressure as a reference. Materials, Methods and Results: After clinical and laboratory evaluation, 34 healthy bitches were selected to undergo elective ovariohysterectomy. After food and water fasting, patients received pethidine hydrochloride intramuscularly as pre-anesthetic medication, followed by anesthetic induction with fentanyl citrate and propofol intravenously. General anesthesia was maintained by inhalation with isoflurane diluted in 100% oxygen. Intraoperative analgesia consisted of continuous infusion of fentanyl citrate intravenously. The animals were randomly divided into 2 groups, the thoracic limb group (TLG) and the pelvic limb group (PLG). In each patient, non-invasive blood pressure measurement was obtained simultaneously with Doppler (DOP) and photoplethysmography (PPG). The sensors of both devices were placed on the end of the same limb. The PPG sensor was positioned in the interdigital region. In patients belonging to the TLG, the Doppler sensor was placed in the ventral region of the thoracic limb, under the ulnar artery. In PLG patients, the Doppler sensor was placed in the dorsal region of the pelvic limb, over the dorsal artery of the foot. The sphygmomanometer was positioned close to the sensors. For systolic blood pressure (SBP) measurement, the cuff was inflated until the Doppler sound signal and the plethysmographic wave were lost. The cuff was then deflated until the Doppler pulse sound resumed and the photoplethysmography showed at least 2 continuous waves on a regular basis. The corresponding pressure value observed on the manometer consisted of the SBP. The same 2 evaluators performed all SBP measurements: 1 responsible for the DOP method and the other for the PPG method; both were blind to the other's findings, thus minimizing potential bias in the results. All animals underwent cannulation of the auricular artery for invasive measurement of systolic blood pressure, using a multiparameter monitor. All blood pressure measurements were performed at 5-min intervals, as well as obtaining additional parameters (heart and respiratory rate, esophageal temperature, partial tissue oxygen saturation, carbon dioxide concentration) and electrocardiographic monitoring. All parameters were documented for further statistical analysis. A strong correlation (r² = 0.95) was obtained between the DOP and PPG methods regardless of the limb on which the sensors were placed. There was a low correlation between the invasive method of measuring systolic blood pressure and the other methods. There was better agreement between the DOP and PPG methods (r2 = -0.0061; P = 0.85) when systolic blood pressure was measured in the TLG. Discussion: In the PLG, the values obtained with the DOP and PPG methods were significantly higher than those obtained with the invasive method, while the values obtained in the TLG differed slightly. It was found that the best measurement site by non-invasive methods was the thoracic limb. It was concluded that the non-invasive methods showed a low correlation with the invasive method; however, both methods had similar characteristics and photoplethysmography can be used to replace the vascular Doppler method.

Polymyositis associated with brainstem signs in a dog

Background: Polymyositis is a generalized inflammatory myopathy which can lead to rhabdomyolysis. This affection may have several origins, including degenerative, metabolic, autoimmune, infectious, inflammatory, ischemic, traumatic, by drug use, induced by toxins and also of idiopathic origin. Diagnosis is made with seric dosage, electrodiagnostic tests and muscle biopsy. Lesions in the rostral oblong medulla may affect the central vestibular system, and there may be signs such as opisthotonos, nystagmus, and strabismus. The aim of this report is to describe a case of a mixed breed dog with manifestation of polymyositis associated with brainstem signs of probable idiopathic origin. Case: A 5-year-old mixed breed male dog was attended with opisthotonos episodes for 2 days, and pelvic limbs extension and thoracic limbs flexion that lasted 10 to 20 min at intervals of approximately 1 h. The animal was anorexic and had also presented one episode of emesis. Upon neurological examination, ventromedial strabismus and Horner's syndrome was observed on the right side, besides vertical nystagmus, flaccid tetraparesis and absence of proprioception in the four limbs. Biochemical analyses revealed creatine kinase (CK) increased (2,433.9 UI/L - reference: 1.5-28.4 UI/L), and urinalysis showed dark color and presence of occult blood without, however, erythrocyturia. Electrocardiogram (ECG) showed QS wave and deviation of the electrical axis. Treatment with prednisolone (1 mg/kg, BID), phenobarbital (2 mg/kg, BID), maropitant citrate (1 mg/kg in 2 doses), and crystalloid fluid therapy (50 mL/kg/day) were prescribed. On the 4th day, the dog was more active and feeding without a tube, so it recommended keep the treatment at home. On the 10th day, the animal had proprioception present on the 4 limbs and normorexia. Biochemical analyses and urinalysis showed no alterations, but normochromic normochromic anemia with thrombocytopenia and leukocytosis by neutrophilia showed in blood count exam. PCR to Ehrlichia canis, Hepatozoon sp., and Babesia canis resulted negative. On the 15th day, blood count, biochemical analyses and urinalysis showed no alterations. Neurological examination revealed only positional vertical nystagmus. which remained as a sequel. Discussion: Polymyositis may be accompanied by rhabdomyolysis, characterized by acute muscle necrosis, increased CK and myoglobinuria. The animal had polymyositis of acute onset, with myoglobinuria and elevated CK values, whose presentation included myalgia and muscle weakness. In humans, polymyositis is accompanied by changes in electrocardiographic tracing without clinical alterations. In dogs, the first report that showed cardiac involvement was compatible with myocarditis. The changes in ECG in the present case was attributed to failure in myocardial electrical conduction. The patient also showed signs of brainstem and central vestibular system injuries. Stress myopathy, intoxication, snakebite, infectious, and metabolic diseases were discarded leading to a clinical suspicion as idiopathic origin. Similar to a published case, the patient of this report received symptomatic and supportive treatment, being discharged from the hospital 20 days after the onset of clinical signs. Thus, polymyositis may be accompanied by signs indicative of brainstem injury. Patients with rhabdomyolysis require intense monitoring due to the high risk of developing acute renal failure. Since no causative agent was identified, symptomatic treatment combined with the prevention of possible complications were fundamental for the maintenance of the animal's life.

Sperm cell capacitation status of ram semen after cooling

Background: The use of conventional artificial insemination (AI) in sheep production is usually associated with lower fertility rates when frozen semen is used. Cooled ram semen has been an alternative over frozen semen due to the higher viability, seminal quality and fertility rates following AI. The semen preservation process promotes sperm cell modifications similar to capacitation (capacitation-like) that causes cell damage affecting viability and seminal quality, but such effects are unclear for cooled semen. The aim of this study was to determine the status of sperm cell capacitation (CA) and acrosome reaction (AR) during ram semen processing and cooling under different extenders, dilution factors, and aerobiosis conditions as a function of storage time at 5o C. Materials, Methods & Results: Two consecutive ejaculates per day per male were collected from 2 adult rams by artificial vagina at 48-72 h intervals, in three replications. After macro- and microscopic evaluations, semen was segregated into groups under 3 extenders (Tris-egg yolk or TY, citrate-egg yolk or CY, skimmed milk or SM), 2 dilution factors (1 x 109 or Bi, 100 x 106 or Mi cells/mL), and 2 aerobiosis conditions (aerobic or A, semi-anaerobic or SA). Diluted semen was cooled to 5ºC and stored for up to 72 h, with evaluations every 24 h. Aliquots of fresh ejaculates and of each cooled diluted subgroup, according to extender, dilution, and aerobiosis, were collected at times T0 and T72 for determination of acrosome status and membrane integrity by the chlortetracycline (CTC) and trypan blue-Giemsa stainings, respectively. No differences were detected in sperm cell motility (M) and motility vigor (V) between fresh and diluted semen. After cooling, a significant decrease in M was observed after 48 h in CY and SM compared with fresh semen and 0 h of cooling, while V started to decrease after 24 h in CY compared with TY. Likewise, M/V from different dilutions and aerobic conditions decreased more significantly after 48 and 24 h of cooling, respectively. The sperm capacitation status did not show differences in the proportion of non-capacitated (NCA), CA and AR sperm cells between TY, CY, and SM extenders (NCA: 75.0%, 71.3%, 74.0%; CA: 15.7%, 17.2%, 15.9%; AR: 9.3%, 11.5%, 10.2%) or between Bi and Mi dilutions (NCA: 74.0%, 72.9%; CA: 15.9%, 16.6%; AR: 10.1%, 10.5%), respectively. However, differences (P < 0.05) were observed between A and SA aerobic conditions, with CA (17.0% vs. 15.5%) and AR (11.9% vs. 8.7%) rates being higher in A than SA, respectively, with no differences in NCA (71.1% vs. 75.8%), irrespective of the storage time. Sperm cell viability decreased after 48 h, especially in CY (P < 0.05). Discussion: Ram sperm cells can suffer irreversible damage due to thermal shock during cooling. Egg yolk-based extenders provide phospholipids and cholesterol to protect the sperm cell membrane during the thermal shock caused by the change in temperature. In this study, sperm cells had irreversible decreases in M/V, with increase in acrosome and plasma membrane damage after cooling to 5ºC. The largest and smallest decreases in M and V over time were observed in the CY and TY extenders, respectively. In addition to the extender type, the semen preservation method and storage time promoted changes in the capacitation status, AR and in sperm cell viability, which per se were associated with a decrease in semen fertility. In fact, the proportions of CA and/or AR sperm cells gradually increased over time after dilution and storage at 5ºC, with a negative correlation between sperm cell viability and M/V over time. In summary, extender and cooling time affected mostly M/V, while aerobiosis condition and dilution factor were more associated with acrosome status and sperm survival, with the extender having less impact on the acrosome status as a function of time.