Utilization of Biotechnology, Neurotransmitter and Cytogenetic Indices in Selecting Pigeon Breeds

Investigations into LDH-A and DR-D4 genes polymorphism, neurotransmitter values and cytogenetic indices of 3 sexed pigeon breeds; non-racing pigeons, (wild rock), racing long distances pigeons (Jan Aarden) and racing short distances pigeons (Janssen) have been performed. The long-distances pigeon showed the highest brain neurotransmitters concentration (p<0.001) among pigeon breeds. Both LDH-A and DR-D4 genes polymorphism indicate the presence of different biodiversity values among pigeon breeds. The variations appeared on the position length 389bp for LDH-A polymorphism, and on two positions length of 418bp and 524bp for DR-D4 polymorphism of long distances male pigeon indicate the presence of unique diversity and overall differences in the amino acids structure in this breed. The protein sequence of both genes showed that in the position of 60 for LDH-A gene the amino acid K (lys) was converted to E (glu), while, in the positions of 117 and 153 for DR-D4 gene the amino acid R (arg) and L (leu) were converted to S (ser) and F (phe) only in long distances male pigeon compared to the other breeds. Moreover, there were slight differences in cytogenetic indices detected among the three pigeon breeds. It can be concluded that both DR-D4 and LDH-A genes polymorphism and neurotransmitters estimations in the brain tissue of racing pigeon would be useful indices for the differentiation and genetic characterization of pigeon breeds and provide a foundation for developing sustainable genetic improvement and conservation programs of the breeding and selecting racing pigeon breeders.(AU)

Hematological parameters in endurance horses pre and post 120 km race / Parâmetros hematológicos em cavalos de enduro pré e pós prova de 120 km

The endurance discipline has the shortest history in Bulgaria compared with other disciplines of equestrian sports. Endurance competitions are held over distances from 40 km to 160 km. The present study focused on the effect of exercise on horses over a distance of 120 km. Changes in the following hematological parameters were investigated:Leukocytes (WBC, g/l); Erythrocytes (RBC, T/l); Platelets (PLT, g/l); Hemoglobin (Hb, g/l); Hematocrit (HCT,%); Mean corpuscular volume(MCV, fl); Mean corpuscular hemoglobin(MCH, pg); Mean cell hemoglobin concentration(MCHC, g/l), Leukogram (Eos; Bas; Neu; Lym; Mon,%) as well as some electrolytes and hormone concentration. The study aimed to establish the changes in these indicators' values before and after the competition. Analysis of the haemogram showed a significant increase in the mean values of 4 of the indicators under study. No significant differences were established in MCV, MCH and MCHC. A decrease was found in the number of Lym, Eos, and Mon and a significant increase in Neu after the competition. The concentration of electrolytes (Ca, Na and K) in the blood serum decreased, and the levels of CK and LDH significantly increased.(AU)

A disciplina de resistência tem a história maiscurta na Bulgária em comparação com outras disciplinas de esportes equestres. As competições de resistência são realizadas em distâncias de 40 km a 160 km. O presente estudo focou no efeito do exercício em cavalos em uma distância de 120 km. Foram investigadas alterações nos seguintes parâmetros hematológicos: Leucócitos (g/l); Eritrócitos (T/l); Plaquetas (g/l); Hemoglobina (g/l); Hematócrito (%); Volume corpuscular médio (VCM, fl); Hemoglobina corpuscular média (HCM, pg); Concentração de hemoglobina corpuscular média(CHCM, g/l), Leucograma (Eos; Bas; Neu; Lin; Mon,%), bem como alguns eletrólitos e concentração hormonal. O estudo teve como objetivo estabelecer as mudanças nos valores desses indicadores antes e depois da competição. A análise do hemograma mostrou um aumento significativo nos valores médios de 4 dos indicadores estudados. Não foram encontradas diferenças significativas no VMC, HCM e CHCM. Foi encontrada uma diminuição no número de Lin, Eos e Mon e um aumento significativo em Neu após a competição. A concentração de eletrólitos (Ca, Na e K) no soro sanguíneo diminuiu e os níveis de Creatina Quinase (CK) e Lactato Desidrogenase (LDH) aumentaram significativamente.(AU)

Pyometra in queens - changes in haemato-biochemical parameters

Background: Pyometra is a bacterial and hormone-induced reproductive disease that occurs in the post-estrus luteal phase in intact queens. Pyometra is more common in the diestrus period due to the high progesterone concentration (in queens that mated, spontaneously ovulated, or were induced to ovulate). However, it can also be seen due to the use of exogenous hormones such as progesterone for the suppression of estrus. More research is needed in cases of pyometra in queens, as well as in bitches. Because, considering that the pathogenesis and characteristics of feline pyometra is similar to bitches, studies on pyometra-affected bitches are taken as reference in studies and applications on queens. From this point of view, the aims of this study were to reveal the changes in complete blood count, blood gas, and serum biochemistry parameters in feline pyometra cases and to determine the correlation between the mentioned parameters. Materials, Methods & Results: In the study, a total of 25 female cats of different breeds were used, between the ages of 6 months and 7 years, 15 were diagnosed with pyometra, and 10 healthy. Anamnesis, clinical findings, and ultrasonographic examinations were used in the diagnosis of pyometra. Abdominal ultrasonography was performed on queens brought to the clinic with complaints such as anorexia, polydipsia, polyuria, abdominal tension, and fever. The control group (n =10) consisted of queens that were introduced to the clinic and were reproductively healthy. Before any treatment in queens with pyometra and the control group, 1 mL blood samples were taken from v. cephalica to evaluate complete blood count, blood gases and serum biochemistry parameters. In complete WBC, Lym, Mon, Gra, RBC, Hb, HCT, MCV, MCH and PLT parameters and, blood gas parameters such as pH, pCO2 , pO2 , sO2 , Na, K, Cl, lactate, glucose, HCO3 , and BE were also evaluated in taken blood samples. Biochemical parameters BUN, creatinine, ALT, AST, ALP, amylase, T.BIL, D.BIL, P, CHOL, TG, LDH, TP, CPK, ALP, Ca, GGT were measured in serum samples. After examination and laboratory analysis, ovariohysterectomy was performed on queens as a treatment. Granulocyte, WBC, HCT and MCH levels of the pyometra group were higher (P < 0.05) and Lym levels were lower (P < 0.05) compared to the control group. According to these results, pH, HCO3 , and BE were lower (P < 0.05) in queens with pyometra than those in the control group, while Na and lactate parameters were higher (P < 0.05). According to the results of biochemical analysis, it was determined that BUN, creatinine levels, GGT, and LDH enzyme activities were found to be higher in the pyometra group compared to the control group, while the Ca level was found to be low (P < 0.05). A positive correlation was observed between BUN and creatinine and LDH, WBC, granulocyte, HCT, and lactate, and a negative correlation between lymphocytes, pH, and BE in the correlation analysis performed on queens with pyometra and control group. However, a positive correlation was observed between creatinine and LDH and HCT, and a negative correlation between lymphocyte, pH and BE. Discussion: There is not enough information about pyometra in queens. As a result, it was determined that there were significant changes in complete blood count, blood gases and serum biochemical parameters in queens with pyometra in this study. These changes were generally thought to be related to dehydration and sepsis or endotoxemia. In addition, it was evaluated that prerenal azotemia occurring in pyometra affected queens may cause renal dysfunction. For this reason, it is thought that the results obtained in the presented study may contribute to the diagnosis, treatment, and prognosis of pyometra cases in queens.

CA 15-3, CRP, and LDH correlates with prognostic parameters in canine mammary neoplasms

The identification of putative prognostic factors in canine mammary neoplasms (CMNs) has been focused on tissue-specific biomarkers, but the serum biomarkers, including cancer antigen 15-3 (CA 15-3), c-reactive protein (CRP), and lactate dehydrogenase (LDH) have been demonstrated to display clinical application in cases of CMNs. The aim of the study was to evaluate the levels of these serum biomarkers and their association with well-established prognostic factors in CMNs. Samples from 15 female canines with CMNs and 15 clinically healthy ones were collected. The results were evaluated using the Tukey's, Pearson, or Spearman tests. The cut-off point, sensitivity, specificity, and area under curve (AUC) were evaluated using the receiver operating characteristic (ROC) curve analysis in a logistic regression model (P<0.05). The levels of CA 15-3, CRP and LDH were significantly higher in the serum of female dogs with CMNs compared to the healthy ones. Moreover, these factors were positively correlated with ulceration, tumor size, histopathological grade, metastatic lymph node, and clinical staging. Female dogs with CMNs were found to exhibit highest serum levels of CA 15-3, CRP, and LDH. Therefore, they can be applied to improve the efficacy of the diagnosis and prognostic evaluation in casas of CMNs.(AU)

Comparative evaluation of plant extract effects on peritoneal, medullary and J774 cells G8 macrophages / Avaliação comparativa do efeito de extratos de plantas em macrófagos peritoneais, medulares e da linhagem J774. G8

The use of medicinal plants as raw material for extracts production and pure substances isolation and subsequence development of new drugs represents a constantly growing area. However, some stages are indispensable before pharmacologically evaluating natural products such as medicines. Toxicity tests in mammalian cells are essential to initiate new drugs development or verify the substance's biocompatibility. Thus, we verified the toxicity of crude extracts and fractions with different polarities obtained from the leaves and stems of eight plant species. The toxic effect was evaluated on macrophages obtained from the bone marrow and peritoneal cavity of a Swiss webster mouse and J774 macrophages. G8 cell lineage. These macrophages were cultured in a 96-well plate, and the compounds were added at a concentration of 100 µg/mL for 24 hours. After this time, the supernatant was removed. The toxicity was evaluated for lactate dehydrogenase (LDH) release assay and the resazurin assay, which uses an indicator dye to measure oxidation-reduction reactions. The results showed a difference in the percentage of toxicity when comparing the same extract in different types of macrophages. This outcome indicates that these cells from different origins may exhibit different responses when exposed to the same natural compounds.

A utilização de plantas medicinais como matéria-prima para a produção de extratos e isolamento de substâncias puras para o desenvolvimento de novos fármacos representa uma área em constante crescimento. No entanto, existem processos a serem realizados antes de avaliar farmacologicamente produtos naturais como medicamentos. Os testes de toxicidade em células de mamíferos são fundamentais para iniciar o desenvolvimento de novas drogas ou verificar a biocompatibilidade de substâncias. Assim, verificamos a toxicidade de extratos brutos e frações com diferentes polaridades obtidos de folhas e caule de oito espécies de planta. Para comparar o efeito tóxico, os testes foram realizados em macrófagos obtidos da medula óssea e cavidade do peritônio de camundongo Swiss webster, bem como no macrófago da linhagem celular J774.G8. Esses macrófagos foram cultivados em placa de 96 poços e os compostos adicionados na concentração de 100 µg/mL por 24 horas. Após esse período o sobrenadante foi removido. A toxicidade foi avaliada pelos ensaios de detecção da enzima lactato-desidrogenase (LDH) e pelo ensaio de resazurina, que usa um corante indicador para medir as reações de oxidação-redução. Os resultados mostraram uma diferença na porcentagem de toxicidade quando comparamos o mesmo extrato em diferentes tipos de macrófagos. Este resultado indica que essas células de várias origens podem exibir respostas distintas quando expostas aos mesmos compostos naturais.

Sheep naturally infected with Anaplasma ovis - evaluation of cardiac and inflammatory biomarkers

Background: Anaplasmosis, also called gall sickness or tropical bovine ehrlichiosis, is an infectious disease caused by species belonging to the genus Anaplasma in domestic and wild animals in tropical and subtropical regions. Anaplasma ovis and A. phagocytophilum are important pathogens of sheep. A. ovis is considered the most common species affecting sheep. The infection is usually subclinical and progresses with high fever, anaemia, icterus, weight loss and abortions. This study aimed to investigate changes in cardiac damage markers, oxidative stress and antioxidant status, cytokines, and acute phase proteins in sheep naturally infected with A. ovis. Materials, Methods & Results: For this purpose, a total of 40 animals, including 20 healthy sheep and 20 sheep infected with anaplasmosis, were used. A. ovis was diagnosed based on clinical findings and peripheral blood smear. Blood smears were prepared from the ear vein. The smears were stained with Giemsa and examined for the presence of Anaplasma spp. Infection was also confirmed by polymerase chain reaction (PCR) analysis. The genomic DNA was isolated from blood, and the MSP-4 gene region was amplified as A. ovis specific target gene. Twenty clinically healthy sheep of the same age group, reared under the same conditions and testing negative in the molecular assessment were used as controls. Blood samples were collected from the cephalic vein and and centrifuged to obtain serum. The serum stored at -20°C until the analysis stage. Serum samples were used for the analysis of cardiac damage markers [troponin I (cTnI), creatine kinase MB (CK-MB), lactate dehydrogenase (LDH) and aspartate transaminase (AST)], oxidative stress parameters [malondialdehyde (MDA), total antioxidant status (TAS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], cytokines [interleukins IL-6, IL-1ß and IL-10, tumour necrosis factor α (TNF-α), and interferon-γ (IFN-γ)] and acute phase proteins [C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp)]. cTnI and CK-MB levels were measured using a chemiluminescent immunoassay. MDA, TAS, SOD, CAT, GPx, TNF-α, IL-1ß, IL-6, IL-10, IFN-γ, SAA and Hp levels were measured by an ELISA reader. LDH, AST and CRP levels were measured in an autoanalyzer. cTnI and LDH levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The concentration of AST was decreased in infected animals. MDA, TAS, SOD, CAT and GPx levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The levels of the inflammatory parameters such as TNF-α, IL-1ß, IL-10 and IFN-γ were significantly increased in the infected animals compared to the healthy ones (P < 0.05). Hp level were significantly increased in the infected group compared with the control group (P < 0.05). However, there was no significant change in CK-MB, SAA and CRP concentrations in the infected animals (P > 0.05). Discussion: Ovine anaplasmosis is an obligate intracellular arthropod disease that causes widespread changes in haematobiochemical, immune response and oxidative stress parameters. Cardiac damage is often overlooked in field conditions due to the lack of adequate knowledge about the pathophysiology of the disease. Our results showed that A. ovis infection leads to significant changes in cardiac biomarkers and that the parasite can cause cardiac dysfunction. This is the first report on cardiac damage markers in Anaplasma-infected sheep. Additionally, the levels of proinflammatory and oxidative stress markers that may cause functional disorders were also found to be increased. Thus, measuring markers of cardiac function, oxidative stress and inflammation can be a useful tool in the early diagnosis of ovine anaplasmosis.

Remote ischemic preconditioning-induced late cardioprotection: possible role of melatonin-mitoKATP-H2S signaling pathway

Purpose: Remote ischemic preconditioning (RIPC) confers cardioprotection against ischemia reperfusion (IR) injury. However, the precise mechanisms involved in RIPC-induced cardioprotection are not fully explored. The present study was aimed to identify the role of melatonin in RIPC-induced late cardioprotective effects in rats and to explore the role of H2 S, TNF-α and mitoKATP in melatoninmediated effects in RIPC. Methods: Wistar rats were subjected to RIPC in which hind limb was subjected to four alternate cycles of ischemia and reperfusion of 5 min duration by using a neonatal blood pressure cuff. After 24 h of RIPC or ramelteon-induced pharmacological preconditioning, hearts were isolated and subjected to IR injury on the Langendorff apparatus. Results: RIPC and ramelteon preconditioning protected the hearts from IR injury and it was assessed by a decrease in LDH-1, cTnT and increase in left ventricular developed pressure (LVDP). RIPC increased the melatonin levels (in plasma), H2 S (in heart) and decreased TNF-α levels. The effects of RIPC were abolished in the presence of melatonin receptor blocker (luzindole), ganglionic blocker (hexamethonium) and mitochondrial KATP blocker (5-hydroxydecanoic acid). Conclusion: RIPC produce delayed cardioprotection against IR injury through the activation of neuronal pathway, which may increase the plasma melatonin levels to activate the cardioprotective signaling pathway involving the opening of mitochondrial KATP channels, decrease in TNF-α production and increase in H2 S levels. Ramelteon-induced pharmacological preconditioning may also activate the cardioprotective signaling pathway involving the opening of mitochondrial KATP channels, decrease in TNF-α production and increase in H2 S levels.

Molecular Investigations of the Effect of Thermal Manipulation During Embryogenesis on the Enzymatic Activity of Creatine Kinase and Lactate Dehydrogenase in Broiler Muscle

Incubating temperature and timing or duration is critical to determine the optimum protocol of thermal manipulation (TM), which underlines muscle growth improvement. Therefore, the aim of the present study is to determine the optimum period of embryonic TM that may result in the improvement of pectoral and thigh muscle myogenesis. This is done by investigating the level of mRNA expression of creatine kinase (CK) and lactate dehydrogenase (LDH). An additional goal is measuring the blood levels of CK and LDH as a biomarker of muscle injury due to the experimental thermal challenge on post-hatch day 35. The study was conducted on 1,440 fertile eggs (Ross broilers) that were divided randomly and equally into a control group and four treatment groups (TM1, TM2, TM3, and TM4). The treatment groups were daily subjected to TM at 39 ºC for 18h with 65% relative humidity (RH) during embryonic days (EDs) 7-11, 11-15, 15-18, and 7-18, respectively. Among the thermally manipulated groups that were investigated, TM1 (ED 7-11) resulted in significant improvement of mRNA expression and enzymatic concentration of CK and LDH in muscle during embryogenesis, as compared to the control. Six hours of TC showed the highest significant CK and LDH expression and concentration levels in the control as compared to TM groups. Thus, the results of this study indicate that TM during ED 7-11 improves pectoral and thigh muscles response to heat stress without adversely affecting their performance. This finding could be used by commercial breeders to enhance local broiler production.(AU)

Molecular Investigations of the Effect of Thermal Manipulation During Embryogenesis on the Enzymatic Activity of Creatine Kinase and Lactate Dehydrogenase in Broiler Muscle

Incubating temperature and timing or duration is critical to determine the optimum protocol of thermal manipulation (TM), which underlines muscle growth improvement. Therefore, the aim of the present study is to determine the optimum period of embryonic TM that may result in the improvement of pectoral and thigh muscle myogenesis. This is done by investigating the level of mRNA expression of creatine kinase (CK) and lactate dehydrogenase (LDH). An additional goal is measuring the blood levels of CK and LDH as a biomarker of muscle injury due to the experimental thermal challenge on post-hatch day 35. The study was conducted on 1,440 fertile eggs (Ross broilers) that were divided randomly and equally into a control group and four treatment groups (TM1, TM2, TM3, and TM4). The treatment groups were daily subjected to TM at 39 ºC for 18h with 65% relative humidity (RH) during embryonic days (EDs) 7-11, 11-15, 15-18, and 7-18, respectively. Among the thermally manipulated groups that were investigated, TM1 (ED 7-11) resulted in significant improvement of mRNA expression and enzymatic concentration of CK and LDH in muscle during embryogenesis, as compared to the control. Six hours of TC showed the highest significant CK and LDH expression and concentration levels in the control as compared to TM groups. Thus, the results of this study indicate that TM during ED 7-11 improves pectoral and thigh muscles’ response to heat stress without adversely affecting their performance. This finding could be used by commercial breeders to enhance local broiler production.

Effect of one-time high load exercise on skeletal muscle injury in rats of different genders: oxidative stress and mitochondrial responses

Purpose: To evaluate the impact of one-time high load exercise on skeletal muscle injury and analysis its mechanism in different genders. Methods: Twenty-four male and 24 female rats were divided randomly into four groups respectively: control, 0 h, 6 h, and 24 h after exercise. The activities of creatine kinase (CK), lactate dehydrogenase (LDH), and myohemoglobin (MYO) in serum, the expression level of oxidative stress markers, mitochondrial respiratory chain complex enzyme, and the apoptosis related protein in quadriceps were detected. Results: The results showed that the activities of CK, LDH and MYO in serum increased immediately after exercise and restored faster in female rats. More obvious structural disorder and apoptosis in male rats were showed. Malondialdehyde (MDA) and superoxide dismutase (SOD) were increased while catalase (CAT) and glutathione (GSH) were decreased in male rats. SOD, CAT and GSH were increased in female rats. Mitochondrial complex enzyme activity was decreased in males and increased in females. Conclusions: The skeletal muscle injury in both genders of rat could be induced by one-time high load exercise due to the mitochondrial respiratory enzyme dysfunction and oxidative stress, which was relatively mild and recovered quicker in female rats.

The effect of size and clinical staging of mammary tumors on blood parameters in bitches

Background: Mammary tumors are the most common type of tumor in female dogs and account for 50% of all tumors in dogs. The clinical prognosis of canine mammary tumors is strongly affected by the size, stages, histological type, and grade of tumor; mitotic index; and nearby and distant metastasis. In canine mammary tumors, it is recommended that prognostic evaluation should also include complete blood count, serum biochemistry, and blood gases in addition to tumor size and stage. This study aimed to investigate the effect of tumor size, volume, and clinical stage on complete blood count, blood gas analysis, and serum biochemical parameters in bitches with mammary tumors and the correlation between them. Materials, Methods & Results: The study included a total of 18 bitches of different breeds, aged 6-15 years, of which 12 had mammary tumors and 6 were healthy. Thoracic X-rays were performed on bitches with mammary tumors in ventrodorsal and laterolateral positions to evaluate lung metastasis. Blood samples were collected from the cephalic vein from bitches in both groups in 2 different tubes (with plastic gel and ethylenediaminetetraacetic acid), 5 mL each, to perform complete blood count and evaluate blood gases and serum biochemical parameters. Blood samples were collected from the animals at the time of initial examination without any intervention. Analysis of the blood showed that bitches with mammary tumors had decreased levels of RBC, HCT, HGB, potassium, TCO2 , base excess, THbc, and ALT enzyme activity and increased levels of lactate, total protein, cholesterol, triglyceride, LDL, uric acid, and ALP and LDH enzyme activities compared with those in the control group. Furthermore, the dogs with a primary tumor of > 5 cm were found to have significantly higher levels of WBC, lactate, total protein, triglyceride, LDL, uric acid, and ALP and LDH enzyme activities and significantly lower levels of RBC and THbc compared with those in the control group. Bitches with tumors in multiple mammary lobes were found to have significantly higher levels of WBC, total protein, triglyceride, LDL, and ALP and LDH enzyme activities and significantly lower levels of RBC, HCT, HGB, TCO2 , THbc, and ALT enzyme activity compared with those in the control group. Based on the laboratory findings and approval of the owners of the dogs, mammary tissues containing the tumor and lymph nodes were surgically removed. After the operation, the removed mammary tissues were evaluated for size and volume. Clinical staging of the tumors was performed based on the size of the primary tumor (T), nearby lymph nodes (N), and metastasis (M) in accordance with the criteria set by WHO. Clinical staging of the tumors was, thus, based on the tumor, nodes, and metastases (TNM) score obtained according to the following system: Stage I: T1 N0 M0 , Stage II: T2 N0 M0 , Stage III: T3 N0 M0 , Stage IV: TanyN1 M0 , Stage V: made as TanyNanyM1 . Discussion: Mammary tumors are the most common type of neoplasm in bitches and, thus, cause serious problems in veterinary medicine. Tumors are significantly correlated with better prognosis compared with larger tumors. Based on this finding, this study investigated the effect of size, volume, and stage of mammary tumors in bitches on some blood parameters and the correlation between them. Therefore, it was concluded that clinical staging and evaluation of blood parameters could be useful in the diagnosis, treatment, and prediction of prognosis in canine mammary tumors. This study found that bitches with mammary tumors exhibited significant changes in their blood parameters (complete blood count, blood gas analysis, and serum biochemistry). The results obtained from this study may contribute to the development of approaches to the diagnosis, prediction of prognosis, and treatment of canine mammary tumors.

Protective effects of melatonin in cisplatin-induced cardiac toxicity: possible role of BDNF-TNF-α signaling pathway

Purpose: The present study explored the role of melatonin in cisplatin-induced cardiac injury along with the possible role of brain-derived neurotrophic factor (BDNF) in melatonin-mediated effects. Methods: Wistar rats were administered cisplatin (10 mg/kg), and cardiac injury was assessed by measuring the levels of cardiac troponin (cTnT) and lactate dehydrogenase (LDH-1).The extent of apoptosis was measured by measuring caspase-3 (pro-apoptotic) and Bcl-2 (anti-apoptotic) in hearts. The levels of BDNF, tumour necrosis factor α (TNF-α) and reduced glutathione were measured in heart. Melatonin (5 and 10 mg/kg) was administered for 15 days, and the role of BDNF was identified by co-administering BDNF inhibitor, ANA-12 (0.25 and 0.5 mg/kg). Results: Melatonin attenuated cTnT and LDH-1 levels along with reduction in caspase-3 and increase in Bcl-2. It also increased cisplatin-induced decrease in BDNF, increase in TNF-α and decrease in reduced glutathione levels. Moreover, ANA-12 abolished the cardioprotective effects, anti-inflammatory and antioxidant effects of melatonin suggesting the role of BDNF in melatonin-mediated effects in cisplatin-induced cardiac injury. Conclusions: Melatonin is useful in cisplatin-induced cardiac injury, which may be due to an increase in BDNF, decrease in inflammation and increase in antioxidant activities.

Pathomorphological and molecular investigations in naturally infected Chukar partridges (Alectoris Chukar) with Histomonas meleagridis and Tetratrichomonas gallinarum / Investigações patomorfológicas e moleculares em perdizes Chukar (Alectoris Chukar) naturalmente infectadas com Histomonas meleagridis e Tetratrichomonas gallinarum

Histomonas meleagridis and Tetratrichomonas gallinarum are two protozoans responsible for mortality associated with typhlohepatitis in poultry. In this study, the etiology of high mortality in Chukar partridges suspected of infection with these agents was investigated pathologically and molecularly. Twelve healthy partridges during the laying period and 30 partridges that died due to disease and were included in the study. In blood analysis, increased levels of WBC, NEU, LYM, MONO, EO and BASO suggesting bacterial and/or parasitic infection; decreased levels of HGB, MCH and MCHC, which are markers of anemia; and increased AST and LDH levels, which are important for liver degenerations. In the liver, which is one of the most pathologically affected organs, multifocal necrosis foci that sometimes merge with each other and spread to large areas, and severe fibrino-necrotic typhlitis were detected. There was amyloid deposition in the space of Disse and vascular sinuses in the liver. PAS positive protozoal agents were observed in and around the lesioned areas. By PCR analyzes using specific primers, 11 of the samples were positive for H. meleagridis only, whereas 5 were positive for T. gallinarum only; 14 samples tested positive for both agents. Sequence analysis showed 100% identity between all samples resulting in positive PCR. In addition, Escherichia coli was produced in microbiological culture (27 of 30). When all the results were evaluated together, it was concluded that H. meleagridis and T. gallinarum and secondary E. coli may cause high mortality in partridges under lay stress.

Histomonas meleagridis e Tetratrichomonas gallinarum são dois protozoários responsáveis pela mortalidade associada à tiflohepatite em aves. Neste estudo, a etiologia da alta mortalidade em perdizes Chukar com suspeita de infecção por esses agentes foi investigada patologicamente e molecularmente. Foram incluídas no estudo 30 perdizes que morreram devido à doença e 12 perdizes saudáveis durante o período de postura. Na análise sanguínea, níveis aumentados de WBC, NEU, LYM, MONO, EO e BASO sugerindo infecção bacteriana e/ou parasitária; diminuição dos níveis de HGB, MCH e MCHC, que são marcadores de anemia; e aumento dos níveis de AST e LDH, que são importantes para as degenerações hepáticas. No fígado, que é um dos órgãos patologicamente mais acometidos, foram detectados focos de necrose multifocais que às vezes se fundem e se espalham para grandes áreas, e tiflite fibrino-necrótica grave no ceco. Houve deposição de amiloide no espaço de Disse e seios vasculares no fígado. Agentes protozoários PAS positivos foram observados dentro e ao redor das áreas lesionadas. Pelas análises de PCR com primers específicos, 11 das amostras foram positivas apenas para H. meleagridis, enquanto 5 foram positivas apenas para T. gallinarum; 14 amostras testaram positivo para ambos os agentes. A análise de sequência mostrou 100% de identidade entre todas as amostras resultando em PCR positivo. Além disso, Escherichia coli foi produzida em cultura microbiológica. Quando todos os resultados foram avaliados em conjunto, concluiu-se que H. meleagridis e T. gallinarum e E. coli secundária podem causar alta mortalidade em perdizes sob estresse de postura.

Stress indicators in cattle submitted to different pre-slaughter rest times / Indicadores de estresse em bovinos submetidos a diferentes tempos de descanso no pré-abate

The objective of this study was to determine the effect of pre-slaughter time about some blood constituents used as stress indicators in the evaluation of animal welfare in cattle. In the practice of ethology, animal welfare is assessed through physiological and behavioral indicators. For the determination of serum biomarkers of animal stress, a total of 180 animals was used, divided into two major groups according to the time of pre-slaughter. In the acceptable group, the animals had a pre-slaughter time up to 24 hours and in the not acceptable group the pre-slaughter time was greater than 24 h. These two groups were split up into three animal categories (males, females and castrated males). In this study, 30 samples were collected from each animal category of the two groups, acceptable and not acceptable, totaling 180 samples. Samples were separated for analysis of muscle enzymes creatine kinase and lactate dehydrogenase (CK and LDH), glucose and cortisol. Physiological values, regardless of the animal category and pre-slaughter time, were higher than the normal values of reference, expressing the prior management effect on animal welfare. The data clearly show a need to update and adapt the entire production chain to animal welfare practices, with the objective of producing competitive quality meat in the world market.(AU)

O objetivo do estudo foi avaliar o efeito do tempo de pré-abate sobre alguns constituintes sanguíneos utilizados como indicadores de estresse na avaliação do bem-estar animal em bovinos. Na prática da etologia, o bem-estar animal é avaliado por meio de indicadores fisiológicos e comportamentais. Para a determinação sérica de biomarcadores de estresse animal utilizaram-se 180 animais, divididos em dois grupos de acordo com o tempo de pré-abate: até 24 horas ou acima de 24 horas. Esses dois grupos subdividiram-se em três categorias animais (machos não-castrados, machos castrados ou fêmeas). Foram coletadas 30 amostras de cada categoria animal nos dois grupos, totalizando 180 amostras. Foram separadas amostras para análise de enzimas musculares creatina quinase e lactato desidrogenase (CK e LDH), glicose e cortisol. Os valores fisiológicos, independentemente da categoria animal e do tempo de pré-abate, foram superiores aos valores de referência, expressando o efeito do manejo prévio sobre o bem-estar dos animais. Os dados evidenciam claramente a necessidade de atualização e adequação de toda a cadeia produtiva às práticas de bem-estar animal, com o objetivo de produzir carne de qualidade competitiva no mercado mundial.(AU)

Babesia bovis in large ruminants in Pakistan - molecular detection and haemato-biochemical alterations

Background: Babesiosis is endemic in Pakistan and is one of the most important bovine diseases that causes huge economic losses and high mortality in young animals. A hematobiochemical study was conducted to unveil the difference between diseased and healthy animals in selected districts i.e., Faisalabad (31° 25' 7.3740'' N and 73° 4' 44.7924'' E), Toba Tek Singh (30° 58' 9.7392'' N and 72° 27' 40.7484'' E) and Jhang (31° 16' 40.9656'' N and 72° 18' 42.3360'' E) of Punjab, Pakistan. Materials, Methods & Results: A total of 518 (Cattle = 360, Buffalo = 158) blood samples were collected. The samples were analyzed by polymerase chain reaction (PCR) targeting apocytochrome b-gene (Babesia bovis-gene) (CYTb) followed by haemato-biochemical analysis. Chi-square test for univariate analysis was used to analyze the data. In summer the PCR-based prevalence was 29.4 (53/180) and 24.05% (19/79) in cows and buffaloes, respectively. On the other hand, in winter results showed that 12.7 (23/180), 13.92 % (11/79) samples positive for Babesia genus from cows and buffaloes, respectively. The positive samples were further investigated for hematological and biochemical analysis. The results revealed that, the mean value of hematological parameters like RBCs, Hb, PCV, MCV and MCHC was significantly (P < 0.05) decreased in infected animals (cows and buffaloes) as compared to the non-infected ones. While the biochemical parameters like Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), cholesterol and Lactate dehydrogenase were significantly (P < 0.05) increased in infected animals as compared to healthy animals. This study is the first molecular and hematobiochemical evidence of Babesia bovis in dairy herds of Punjab province, Pakistan. Discussion: Bovine babesiosis is one of the important tick-borne diseases (TBD) affecting dairy industry. In bovines, among 3 Babesia species that cause the disease B. bovis is more pathogenic with high mortality and morbidity. Pakistan is situated in tropical and sub-tropical region where the humidity is high in some part of countries. This high humidity mostly favors the reproduction of the ticks thus higher prevalence of TBDs in this region. Initially the babesiosis was diagnosed by light microscopy using thin blood smear stained with Giemsa stain. Many studies verified that PCR is a more specific and sensitive tool than conventional techniques for the detection of carrier / asymptomatic ruminants. The haemato-biochemical profile is another valuable footprint to track the disease. Keeping in view the above-mentioned fact the present project has been planned to evaluate the haemato-biochemical alteration between health and Babesia infected cattle along with the molecular detection of Babesia species involved in bovine babesiosis. The mean values of haematobiochemical parameters in clinically ill and healthy animals revealed that the mean values of hematological parameters like RBCs, Hb, PCV, and HCT were significantly decreased in diseased animals as compared to the healthy ones. All these might be due the fact that the parasite is intra-erythrocytic in nature and destruction of red blood cells results in significant (P < 0.05) decrease level of all the hematological parameters. The mean value of ALT in babesiosis infected cattle was significantly higher as compared to healthy cattle. The mean values of AST and LDH in babesiosis infected cows was significantly higher as compared to that in healthy cows. The elevation in liver enzymes in babesiosis may be due to the hepatic damage and lesions induced by the parasite during its multiplication in the blood followed by disturbed liver function. These enzymes are present in high concentrations in the muscles and liver. High level of these enzymes in the blood is indicator of organ necrosis or damage.

Enzymatic markers of pulmonary inflammation in tracheal wash fluid in racehorses: alkaline phosphatase and lactate dehydrogenase

Background: Diagnosis of the lower airway diseases (LAD) in horses relies on clinical signs, endoscopic examinationand cytologic evaluation of lower respiratory tract flushing fluids as well. Specific enzyme activities in respiratory tractfluids of horses could be used as useful indicators in lower airway disorders. The main goal of the study was to evaluate thediagnostic usefulness of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) enzyme activites in tracheal wash(TW) fluids of horses with lower respiratory tract inflammation and/or damage and also in distinguishing of non-septicand septic inflammation in racehorses with LAD.Materials, Methods & Results: A cross-sectional study was conducted in racehorses. TW fluid samples were obtainedfrom 83 racehorses (71 with lower airway diseases and 12 healthy controls) for the study. The One-Way ANOVATest was used for the significance analysis of the differences between the groups in showing the normal distributionof blood serum ALP and LDH properties. Kruskal Wallis H Test and Tamhane T2 test were used for the significanceanalysis of the differences between the groups of tracheal ALP and LDH properties, which were not normally distributed. The sampled horses were classified into 3 groups as healthy control, non septic neutrophilic inflammation(NS-LAD) and septic neutrophilic inflammation (S-LAD) according to the tracheal mucus scores determined duringendoscopic examination, clinical signs, cytologic and bacteriologic examination of collected TW samples. Accordingto cytology results normal neutrophil levels (< 20%) were determined in the healthy control group (14.5%). However, increased neutrophil levels (≥ 20% and ≥ 80%) were encountered in the NS-LAD (61.4%) and S-LAD (24.1%)groups respectively. Also, the signs of degenerative changes and presence of intracellular bacteria in the neutrophilcells in the S-LAD group were observed. Bacteriologic analysis of the samples...

Enzymatic markers of pulmonary inflammation in tracheal wash fluid in racehorses: alkaline phosphatase and lactate dehydrogenase

Background: Diagnosis of the lower airway diseases (LAD) in horses relies on clinical signs, endoscopic examinationand cytologic evaluation of lower respiratory tract flushing fluids as well. Specific enzyme activities in respiratory tractfluids of horses could be used as useful indicators in lower airway disorders. The main goal of the study was to evaluate thediagnostic usefulness of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) enzyme activites in tracheal wash(TW) fluids of horses with lower respiratory tract inflammation and/or damage and also in distinguishing of non-septicand septic inflammation in racehorses with LAD.Materials, Methods & Results: A cross-sectional study was conducted in racehorses. TW fluid samples were obtainedfrom 83 racehorses (71 with lower airway diseases and 12 healthy controls) for the study. The One-Way ANOVATest was used for the significance analysis of the differences between the groups in showing the normal distributionof blood serum ALP and LDH properties. Kruskal Wallis H Test and Tamhane T2 test were used for the significanceanalysis of the differences between the groups of tracheal ALP and LDH properties, which were not normally distributed. The sampled horses were classified into 3 groups as healthy control, non septic neutrophilic inflammation(NS-LAD) and septic neutrophilic inflammation (S-LAD) according to the tracheal mucus scores determined duringendoscopic examination, clinical signs, cytologic and bacteriologic examination of collected TW samples. Accordingto cytology results normal neutrophil levels (< 20%) were determined in the healthy control group (14.5%). However, increased neutrophil levels (≥ 20% and ≥ 80%) were encountered in the NS-LAD (61.4%) and S-LAD (24.1%)groups respectively. Also, the signs of degenerative changes and presence of intracellular bacteria in the neutrophilcells in the S-LAD group were observed. Bacteriologic analysis of the samples...(AU)

Clinical and laboratory findings in sheep experimentally poisoned by Palicourea marcgravii and the use of magnesium chloride with possible therapeutic effect on poisoning

ABSTRACT: This study aimed to examine the efficacy of magnesium chloride therapy in sheep experimentally poisoned by Palicourea marcgravii and describe the clinical and laboratory findings that may aid in early ante mortem diagnosis of this poisoning. P. marcgravii is an important plant within a group of 22 species in Brazil that cause sudden death. Its toxic component is sodium monofluoroacetate. It was observed that a time for evolution of poisoning was longer in our study compared to other studies that used the same dose of P. marcgravii in sheep. However, all animals that were poisoned and subsequently treated with magnesium chloride died. The sheep presented characteristic clinical signs before death in addition to other signs that have not been previously described for this type of poisoning, such as abdominal breathing, coughing, head pressing, and nystagmus. This is the first evaluation of hematological parameters of sheep poisoned by P. marcgravii. Leukocytosis with neutrophilia, hyperfibrinogenemia, and hyperglycemia, associated with hypophosphatemia and elevated levels of serum aspartato aminotransferase (AST), glutamate dehydrogenase (GLDH), urea, creatinine, creatine kinase (CK) and lactate dehydrogenase (LDH), were observed. Additionally, changes were noted in necropsy, such as engorged jugular, large vessel congestion, pulmonary edema. Histological examination revealed vacuolar hydropic degeneration in the distal twisted uriniferous tubules in the kidneys, similar to those reported in previous studies on P. marcgravii poisoning.

RESUMO: O presente estudo objetivou verificar a eficácia terapêutica do cloreto de magnésio em ovinos intoxicados experimentalmente por Palicourea marcgravii e descrever os achados clínicos e laboratoriais que possam auxiliar no diagnóstico precoce ante mortem dessa intoxicação. P. marcgravii é a principal planta dentro de um grupo de 22 espécies que causam morte súbita no Brasil e seu princípio tóxico é o monofluoracetato de sódio. Durante o procedimento, observou-se maior tempo de evolução em relação a outros trabalhos utilizando-se a mesma dose da P. marcgravii em ovinos, entretanto todos os animais intoxicados e posteriormente tratados com cloreto de magnésio vieram a óbito. Os ovinos apresentaram sinais clínicos característicos antes da morte, associados a sinais nunca antes descritos nesse tipo de intoxicação, como respiração abdominal, tosse, head pressing e nistagmo. Trata-se da primeira avaliação dos parâmetros hematológicos de ovinos intoxicados por P. marcgravi. Foi observado leucocitose com neutrofilia, hiperfibrinogemia e hiperglicemia, associado a elevação dos valores séricos de AST, GLDH, ureia, creatinina, CK, LDH e hipofosfatemia. À necropsia foram notadas alterações como jugular ingurgitada, congestão de grandes vasos, edema pulmonar. Histologicamente havia degeneração hidrópica vacuolar nos túbulos uriníferos contorcidos distais no rim, semelhantes às já relatadas em estudos anteriores, na intoxicação por P. marcgravii.

Anticoccidial and Biochemical Effects of Artemisia Brevifolia Extract in Broiler Chickens

This study was conducted to evaluate the anticoccidial potential of Artemisia brevifolia extract (ABE) and its effect on biochemical parameters against experimental coccidiosis in broiler chickens. For this purpose, a total of 150 broiler chicks were procured and divided into 5 equal groups (30 birds per group). At one week of age, the first three groups (A, B and C) were orally fed with ABE at dose rates of 100, 200 and 300mg/kg respectively for seven consecutive days. Group D served as infected/medicated and was treated with Baycox® (1ml/L of water). Group E served as the infected/non-medicated control group. At two weeks of age, all groups were infected with 50,000 sporulated oocysts of Eimeria tenella. The anticoccidial potential was evaluated based on lesion score, oocyst score, fecal score, feed conversion ratio, and mortality (%). Furthermore, the effect of ABE on hematological parameters and serum chemistry was also evaluated by following standard procedures. The anticoccidial effects were compared with the standard drug Baycox® (p>0.05). Results of the study showed the anticoccidial activity of ABE in terms of lesion diminishing, oocyst and fecal scores, better feed conversion ratios, and mortality reduction (%). The ABE also improved hematological parameters and stabilized the serum enzymes (ALT, LDH, Creatinine) of infected chicks (p>0.05).(AU)

Anticoccidial and Biochemical Effects of Artemisia Brevifolia Extract in Broiler Chickens

This study was conducted to evaluate the anticoccidial potential of Artemisia brevifolia extract (ABE) and its effect on biochemical parameters against experimental coccidiosis in broiler chickens. For this purpose, a total of 150 broiler chicks were procured and divided into 5 equal groups (30 birds per group). At one week of age, the first three groups (A, B and C) were orally fed with ABE at dose rates of 100, 200 and 300mg/kg respectively for seven consecutive days. Group D served as infected/medicated and was treated with Baycox® (1ml/L of water). Group E served as the infected/non-medicated control group. At two weeks of age, all groups were infected with 50,000 sporulated oocysts of Eimeria tenella. The anticoccidial potential was evaluated based on lesion score, oocyst score, fecal score, feed conversion ratio, and mortality (%). Furthermore, the effect of ABE on hematological parameters and serum chemistry was also evaluated by following standard procedures. The anticoccidial effects were compared with the standard drug Baycox® (p>0.05). Results of the study showed the anticoccidial activity of ABE in terms of lesion diminishing, oocyst and fecal scores, better feed conversion ratios, and mortality reduction (%). The ABE also improved hematological parameters and stabilized the serum enzymes (ALT, LDH, Creatinine) of infected chicks (p>0.05).