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)

Effect of Bacillus Subtilis on Immune Function of Hd11 Chicken Macrophages

Poultry is frequently contaminated by Salmonella, a pathogen leading to human health concern worldwide. This study aimed to evaluate the effect of Bacillus subtilis (BS)strain 048 (BS048) on the activation, phagocytosis, sterilization, cytokine secretion, and nitrogen oxide synthesis of HD11 chicken macrophages subjected to Salmonella enteritidis challenge, using lipopolysaccharide treatment as a negative control. The results showed: (1) BS048 had no significant effect on extracellular lactate dehydrogenase activity (p>0.05), while lipopolysaccharide treatment significantly increased extracellular lactate dehydrogenase activity (p >0.05), while lipopolysaccharide treatment significantly increased extracellular lactate dehydrogenase activity (p 0.05);(2)BS048 significantly upregulated the expression levels of pro-inflammatory cytokines (interleukin (IL)- 1â and IL-6), anti-inflammatory cytokines(IL-10 and transforming growth factor-â1), and anti-viral cytokine, interferon-â (p<0.01); ; (3) BS048 significantly upregulated the mRNA expression level of the inducible nitric oxide synthase and its activity as well as extracellular nitrogen oxide level (p <0.01). In conclusion, BS048 could improve antiinflammatory and immune functions of HD11 chicken macrophages, without cytotoxic effects on these cells.(AU)

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)

Effect of monochrome light with different wavelengths on biochemical parameters of hens

Artificial light, as one of the environmental factors, plays a significant role in regulating the synthesis and secretion of hormones related to the coordination of parameters of life, growth, immunity, and reproductive functions of hens. The article aims to study the influence of monochrome light with different wavelengths on the biochemical parameters of hens` blood serum. Four groups of "Hy-Line W-36" crossbred hens were formed. Hens of the 1st group were kept using monochrome light with different wavelength lamps with a wavelength of ~ 460 nm, the 2nd group ~ 600 nm, the 3rd group ~ 630 nm, and the 4th group ~ 650 nm. It was found that the use of light with different wavelengths for keeping hens in cages of multilevel batteries affects hen' biochemical parameters, according to the research results. It was established that when using light with a wavelength of ~ 630 and ~ 650 nm, the indicators of clinical biochemistry of hens' blood serum were within the normal physiological values. Whereas, with the use of light with a wavelength of ~ 600 nm, an increase in the level of glucose, creatinine, total protein, total bilirubin, and phosphorus, a decrease in the ratio of calcium to phosphorus, in the activity of alkaline phosphatase, aspartate aminotransferase, and lactate dehydrogenase, were observed in the hens' blood serum. The use of light with a wavelength of ~ 460 nm was accompanied by a further increase in the level of glucose, creatinine, total protein, urea, total bilirubin, phosphorus, the activity of alkaline phosphatase, aspartate aminotransferase, lactate dehydrogenase, and gamma-glutamyltransferase, a decrease in the ratio of calcium and phosphorus.

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.

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.

Poliartrite em cão por Leishmaniose - tratamento com miltefosina / Polyarthritis in a dog due to leishmaniasis - treatment with miltefosine

Background: Canine visceral leishmaniasis causes several clinical signs, such as lymphadenomegaly, exfoliative dermatitis, ulcerative skin lesions, and lameness. The most commonly reported locomotor changes are claudication, edema, arthralgia, joint stiffness, and muscle atrophy. Radiographic exam revealed cortical and medullary destruction, increase, or decrease in medullary opacity, proliferative periosteal reaction, osteolysis, collapse of joint spaces and soft tissue edema are observed. The aim of this report is to describe the clinical and radiographic evolution of a case of erosive polyarthritis associated with leishmaniasis in a dog before, during and after treatment with miltefosine. Case: A 7-month-old mixed-breed dog was attended due pain and limited mobility. In the orthopedic evaluation, joint swelling, stiffness, and increased pain sensitivity of the four limbs, as well as neck stiffness, were noted. Radiographic examination showed joint changes compatible with edema, with increased volume and radiopacity of the soft tissues adjacent to the joints. The segments of the patient's spine showed more severe bone alterations, the cervical spine being one of the most affected regions, with multiple bone proliferations throughout the vertebral body, especially in the ventral portion (spondylosis), compatible with polyarthritis due to leishmaniasis. Due to the suspicion, lymph node and spleen cytology was performed, confirming the diagnosis. Hematological examination revealed anemia, leukopenia due to lymphopenia and thrombocytopenia in addition to increased AST (79,4 U/L; reference: 6,2 - 13 U/L), creatine kinase (517,6 U/L; reference: 1,5 - 28,4 U/L), lactate dehydrogenase (688,4 IU/L; reference: 45 - 233 IU/L) and hyperproteinemia (7,34 g/dL; reference: 5,4 - 7,1 g/dL). Treatment with miltefosine, allopurinol, domperidone, prednisone, gabapentin and dipyrone was started. Reassessments were performed monthly for 3 consecutive months. Hematological examinations showed improvement, with resolution of anemia and thrombocytopenia, and a marked decrease in creatine kinase values. Thus, it is evident that the dog did not develop liver or kidney changes during treatment. During the treatment and monitoring in this period, the dog had a clinical improvement, which started to walk without pain. In addition, joint swellings were no longer present, however, there was no improvement in the radiographic evaluation of the joints. Discussion: Clinical signs of the locomotor system are compatible with those described in animals that had osteoarticular manifestations associated with leishmaniasis, such as arthralgia, edema, and joint stiffness. In the present report, treatment with miltefosine associated with allopurinol resulted in an improvement in the clinical picture, and this therapy is therefore promising in dogs with polyarthritis due to leishmaniasis. A case published in human medicine demonstrated the intra-articular absorption capacity of this drug. There is only one study to date that describes the radiographic evolution of a dog with arthritis due to leishmaniasis after treatment with miltefosine and allopurinol. In this case described, the dog reported remained with the osteoarticular lesions after treatment, although clinical improvement was observed, as in our report. The use of miltefosine and allopurinol are in accordance with stage II staging for leishmaniasis. In this study, although there was no improvement in the radiographic examinations, the treatment was effective in the remission of the animal's clinical condition.

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.

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.

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.

In vitro antibacterial activities of silver nanoparticles synthesised using the seed extracts of three varieties of Phoenix dactylifera

Abstract Green synthesis of silver nanoparticles (AgNPs) is an ecofriendly, cost-effective and promising approach for discovery of novel therapeutics. The aim of the current work was to biogenic synthesize, characterize AgNPs using seed extracts of three economically important varieties of date palm (Iklas, Irziz and Shishi), and assess their anti-pathogenic bacterial activities. AgNPs were synthesised then characterised using electron microscopy and Fourier transform infrared analyses. The bactericidal activities of AgNPs against five different bacterial pathogens, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus and Streptococcus pneumoniae, were determined in vitro. In particular, changes in membrane integrity of virulent bacterial strains in response to AgNPs were investigated. Results of lactate dehydrogenase, alkaline phosphatase activity assays, and measurement of membrane potential revealed that the cytotoxic effects of the AgNPs were mainly centred on the plasma membrane of bacterial cells, leading to loss of its integrity and eventually cell death. In conclusion, green synthesis of AgNPs is an efficient, cost-effective and promising strategy to combat virulent antibiotic-resistant strains.

Resumo A síntese verde de nanopartículas de prata (AgNPs) é uma abordagem ecologicamente correta, econômica e promissora para a descoberta de novas terapêuticas. O objetivo do presente trabalho foi sintetizar biogênica, caracterizar AgNPs usando extratos de sementes de três variedades economicamente importantes de tamareira (Iklas, Irziz e Shishi) e avaliar suas atividades bacterianas antipatogênicas. AgNPs foram sintetizados e caracterizados usando microscopia eletrônica e análise de infravermelho por transformada de Fourier. As atividades bactericidas de AgNPs contra cinco diferentes patógenos bacterianos, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Staphylococcus aureus resistente à meticilina e Streptococcus pneumoniae, foram determinadas in vitro. Em particular, foram investigadas alterações na integridade da membrana de cepas bacterianas virulentas em resposta a AgNPs. Os resultados da lactato desidrogenase, dos ensaios da atividade da fosfatase alcalina e da medição do potencial de membrana revelaram que os efeitos citotóxicos dos AgNPs estavam principalmente centrados na membrana plasmática das células bacterianas, levando à perda de sua integridade e, eventualmente, à morte celular. A síntese verde de AgNPs é uma estratégia eficiente, econômica e promissora para combater cepas virulentas resistentes a antibióticos.

Assessment of a vaquejada horse training protocol based on laboratory clinical parameters

The objective of this study was to assess a training protocol employed in the Brazilian Northeast region for fitness conditioning of vaquejada horses. For 12 months, 24 Quarter Horses were evaluated under a completely randomized split-plot experimental design in which the plots comprised three age groups: horses at two, three, and four years of age. The split plots were made up of six fitness tests carried out every other month. The fitness test protocol consisted of five levels of protocol exercises on a standard vaquejada track. Prior to the tests with fasted animals, we collected blood samples to determine muscle enzymes (aspartate aminotransferase, creatine kinase, and lactate dehydrogenase). During the tests, heart rate, speed, and distance run were recorded using a heart rate monitor. Next, the results were used to calculate speed at which each horse reached 150 bpm (V150), speed at which each horse reached 200 bpm (V200), maximum heart rate (HRmax), maximum speed (Vmax), recovery time needed for the heart rate of horses to return to half the maximum value reached during the fitness tests (HR50%), and recovery time needed for the heart rate of horses to return to baseline values (HRbasal). No difference was found among the age groups for V150, HRmax, Vmax, HR50%, HRbasal, or muscle enzymes. By the final stage of training, the V200 of the three-year-old horses was higher than that of the four-year-old foals. During training, all groups exhibited increases in serum concentrations of muscle enzymes and reductions in efficiency to recover heart rate after exercise. The training protocol assessed is unable to maintain proper fitness for competitions throughout the year.(AU)

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.

Cardioprotective action of aprepitant in a rat model of ischemia-reperfusioninduced myocardial injury: role of PI3K-AkT-GSK-3ß-HIF-1α signaling pathway

Purpose: The present study explored the role and mechanism involved in aprepitant-induced cardioprotective effects in rat model of ischemia-reperfusion injury. Methods: The isolated hearts of Wistar male albino rats were subjected to ischemia-reperfusion injury on Langendorff apparatus. The extent of myocardial injury was assessed by measuring lactate dehydrogenase 1 and CK-MB release in the coronary effluent. The rats were treated with aprepitant (5, 10 and 20 mg/kg) before isolating hearts. After injury, the levels of HIF-1α, p-AkT, p-GSK-3ß/GSK-3ß were measured in heart homogenates. LY294002 was employed as PI3K inhibitor. Results: Ischemia-reperfusion led to significant myocardial injury and decreased the levels of HIF-1α, p-AkT and ratio of p-GSK-3ß/GSK-3ß. Aprepitant attenuated myocardial injury and restored the biochemical changes in a dose-dependent manner. Pre-treatment with LY294002 (10 and 20 mg/kg) abolished aprepitant-mediated cardioprotective effects and restored the biochemical parameters in the heart homogenate. Conclusions: Aprepitant may be effective in preventing ischemia-reperfusion-induced myocardial injury, which may be due to activation of PI3K-AkT-GSK-3ß and HIF-1α signaling pathway.

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.

In vitro antibacterial activities of silver nanoparticles synthesised using the seed extracts of three varieties of Phoenix dactylifera / Atividades antibacterianas in vitro de nanopartículas de prata sintetizadas usando extratos de sementes de três variedades de Phoenix dactylifera

Green synthesis of silver nanoparticles (AgNPs) is an ecofriendly, cost-effective and promising approach for discovery of novel therapeutics. The aim of the current work was to biogenic synthesize, characterize AgNPs using seed extracts of three economically important varieties of date palm (Iklas, Irziz and Shishi), and assess their anti-pathogenic bacterial activities. AgNPs were synthesised then characterised using electron microscopy and Fourier transform infrared analyses. The bactericidal activities of AgNPs against five different bacterial pathogens, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus and Streptococcus pneumoniae, were determined in vitro. In particular, changes in membrane integrity of virulent bacterial strains in response to AgNPs were investigated. Results of lactate dehydrogenase, alkaline phosphatase activity assays, and measurement of membrane potential revealed that the cytotoxic effects of the AgNPs were mainly centred on the plasma membrane of bacterial cells, leading to loss of its integrity and eventually cell death. In conclusion, green synthesis of AgNPs is an efficient, cost-effective and promising strategy to combat virulent antibiotic-resistant strains.

A síntese verde de nanopartículas de prata (AgNPs) é uma abordagem ecologicamente correta, econômica e promissora para a descoberta de novas terapêuticas. O objetivo do presente trabalho foi sintetizar biogênica, caracterizar AgNPs usando extratos de sementes de três variedades economicamente importantes de tamareira (Iklas, Irziz e Shishi) e avaliar suas atividades bacterianas antipatogênicas. AgNPs foram sintetizados e caracterizados usando microscopia eletrônica e análise de infravermelho por transformada de Fourier. As atividades bactericidas de AgNPs contra cinco diferentes patógenos bacterianos, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Staphylococcus aureus resistente à meticilina e Streptococcus pneumoniae, foram determinadas in vitro. Em particular, foram investigadas alterações na integridade da membrana de cepas bacterianas virulentas em resposta a AgNPs. Os resultados da lactato desidrogenase, dos ensaios da atividade da fosfatase alcalina e da medição do potencial de membrana revelaram que os efeitos citotóxicos dos AgNPs estavam principalmente centrados na membrana plasmática das células bacterianas, levando à perda de sua integridade e, eventualmente, à morte celular. A síntese verde de AgNPs é uma estratégia eficiente, econômica e promissora para combater cepas virulentas resistentes a antibióticos.

Influence of bentonite adsorbents associated with hepatic antioxidants on the health and production of dairy cows fed diets containing naturally produced mycotoxins / Influência do adsorvente bentonita associado a antioxidantes hepáticos na saúde e produção de vacas leiteiras alimentadas com dietas contendo micotoxinas naturalmente produzidas

As mycotoxins are consistent contaminants in the dairy cow diet, the use of adsorbents is recommended, although there are no ideal adsorbents. Although there are studies on this subject, few have focused on chronic natural intoxications. Here, we evaluated the effect of bentonite adsorbents associated with liver antioxidants on the health and milk production of dairy cows fed a diet containing naturally-produced fumonisin, zearalenone, and deoxynivalenol. Eighteen dairy cows (bodyweight 550 ±50 kg, 5 ± 2 years old, and 30 ± 1,25 kg/day milk production) in the middle of lactation were divided into groups: treatment (TG, n = 9, 22 g/day of supplement added to diet) and control (CG, n = 9, without supplement). A physical examination was performed, weekly over 56 days and blood was collected for liver and immune assessments. Milk was harvested to evaluate milk production and content (fat protein, somatic cell count, and lactose). The additive promoted beneficial effects on the liver from the 24th day due to a decrease in the enzymatic activities of gamma-glutamyltransferase and lactate dehydrogenase and increased serum protein and albumin levels. There were improvements in health, evidenced as fewer clinical manifestations of the disease, greater leukocyte oxidative metabolism capacity, and a lower neutrophil lymphocytes ratio. The treatment also promoted a 19% increase in milk volume. It was concluded that the additive promoted health benefits and milk production in dairy cows.(AU)

Como as micotoxinas são contaminantes constantes na dieta de vacas leiteiras, o uso de adsorventes é recomendado, embora não existam adsorventes ideiais. Ainda que existam estudos sobre o assunto, poucos são direcionados às intoxicações naturais crônicas. Nosso experimento avaliou o efeito do adsorvente bentonita associado a antioxidantes hepáticos sobre a saúde e produção de leite de vacas leiteiras alimentadas com dieta contendo fumonisina, zearalenona e desoxinivalenol produzidos naturalmente. Dezoito vacas lei-teiras (peso corporal 550 ±50 kg, 5±2 anos de idade, e 30 ± 1,25 kg/dia de produção de leite) no meio da lactação, foram divididas em grupos: Tratamento (GT, n=9, 22g/dia do suplemento adicionado à dieta) e controle (GC, n =9, sem suplemento). Em intervalos semanais, durante 56 dias, foi realizado exame físico, coleta de sangue para avaliação hepática e imunológica. O leite foi colhido para avaliar a produção e análises de leite para (proteína, gordura, contagem de células somáticas e lactose). O aditivo promoveu efeitos bené-ficos no fígado à partir do 24º dia devido à diminuição das atividades enzimáticas da gama-glutamiltransferase e lactato desidrogenase e aumento dos níveis séricos de proteína e albumina. Houve melhora na saúde, evidenciada por menor número de manifestações clínicas de doença, maior capacidade de metabolismo oxidativo dos leucócitos e menor razão neutrófilos/linfócitos. O tratamento também promo-veu aumento de 19% no volume de leite. Concluiu-se que o aditivo promoveu benefícios à saúde e à produção de leite em vacas leiteiras.(AU)

In vitro antibacterial activities of silver nanoparticles synthesised using the seed extracts of three varieties of Phoenix dactylifera / Atividades antibacterianas in vitro de nanopartículas de prata sintetizadas usando extratos de sementes de três variedades de Phoenix dactylifera

Green synthesis of silver nanoparticles (AgNPs) is an ecofriendly, cost-effective and promising approach for discovery of novel therapeutics. The aim of the current work was to biogenic synthesize, characterize AgNPs using seed extracts of three economically important varieties of date palm (Iklas, Irziz and Shishi), and assess their anti-pathogenic bacterial activities. AgNPs were synthesised then characterised using electron microscopy and Fourier transform infrared analyses. The bactericidal activities of AgNPs against five different bacterial pathogens, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus and Streptococcus pneumoniae, were determined in vitro. In particular, changes in membrane integrity of virulent bacterial strains in response to AgNPs were investigated. Results of lactate dehydrogenase, alkaline phosphatase activity assays, and measurement of membrane potential revealed that the cytotoxic effects of the AgNPs were mainly centred on the plasma membrane of bacterial cells, leading to loss of its integrity and eventually cell death. In conclusion, green synthesis of AgNPs is an efficient, cost-effective and promising strategy to combat virulent antibiotic-resistant strains.

A síntese verde de nanopartículas de prata (AgNPs) é uma abordagem ecologicamente correta, econômica e promissora para a descoberta de novas terapêuticas. O objetivo do presente trabalho foi sintetizar biogênica, caracterizar AgNPs usando extratos de sementes de três variedades economicamente importantes de tamareira (Iklas, Irziz e Shishi) e avaliar suas atividades bacterianas antipatogênicas. AgNPs foram sintetizados e caracterizados usando microscopia eletrônica e análise de infravermelho por transformada de Fourier. As atividades bactericidas de AgNPs contra cinco diferentes patógenos bacterianos, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Staphylococcus aureus resistente à meticilina e Streptococcus pneumoniae, foram determinadas in vitro. Em particular, foram investigadas alterações na integridade da membrana de cepas bacterianas virulentas em resposta a AgNPs. Os resultados da lactato desidrogenase, dos ensaios da atividade da fosfatase alcalina e da medição do potencial de membrana revelaram que os efeitos citotóxicos dos AgNPs estavam principalmente centrados na membrana plasmática das células bacterianas, levando à perda de sua integridade e, eventualmente, à morte celular. A síntese verde de AgNPs é uma estratégia eficiente, econômica e promissora para combater cepas virulentas resistentes a antibióticos.