Leech therapy (Hirudo medicinalis) attenuates testicular damages induced by testicular ischemia/reperfusion in an animal model.

BACKGROUND: Testicular torsion/detorsion triggers tissue ischemia/reperfusion, leading to reactive oxygen species overgeneration and apoptosis. The saliva of leeches is full of anti-inflammatory, anticoagulants, antioxidants, and antimicrobial agents. Therefore, this study aimed to assess the protective mechanism of leech therapy on testicular ischemia/reperfusion damage. METHODS: 18 adult male rats were randomly divided into three groups: 1-Sham-operated group (SO). 2-Torsion/detorsion (T.D) group: two hours of testicular torsion with two hours of testicular detorsion was performed. 3-Torsion/detorsion + Leech therapy (TDL) group. Sperm parameters (motility, vitality, morphology, and concentration), oxidative stress biomarkers (MDA, CAT, GPx, and TAC), histopathological factors (Mean seminiferous tubular diameter, Germinal epithelial cell thickness, Testicular capsule thickness, Johnson's score, and Cosentino's score), and immunohistochemical markers for apoptosis detection (Bax, Bcl-2, and Caspase-3) were measured. RESULTS: There was a significant difference for all sperm parameters in the T. D group compared to the sham group. Leech therapy significantly increased progressive motility and normal morphology and reduced non-progressive motility. In the TDL group, MDA concentration significantly reduced, and levels of GPx, TAC, and CAT remarkably increased. All evaluated histopathological parameters in the TDL group significantly increased compared to the T. D group except for the testicular capsule thickness. T. D notably increased the expression of Bax and Caspase-3, while the treatment group slowed the rate of apoptosis compared to the control group. Bcl-2 expression in the T. D group was significantly lower than that in the sham group. Leech therapy increased the Bcl-2 expression. CONCLUSION: Leech therapy attenuates damages to testicular tissue following torsion/detorsion due to its antioxidant, anti-inflammatory, and anti-apoptotic effects. Hence, it can be considered as an effective remedy for testicular ischemia/reperfusion.

Co-administration of Salvia miltiorrhiza and verapamil inhibits detrimental effects of torsion/detorsion on testicular tissue in rats.

Testicular torsion/detorsion is one of the important emergencies that requires fast surgical intervention. This study aimed to investigate the effects of Salvia miltiorrhiza hydroalcoholic extract combined with verapamil on testicular ischaemia/reperfusion damage in Wistar albino rats. All animals were distributed in 3 groups (n = 8), including the sham-operated group, torsion/detorsion (TD) group and torsion/detorsion + pretreatment with 200 mg/kg Salvia miltiorrhiza extract combined with 0.3 mg/kg verapamil (SMV) group. Oxidative stress biomarkers (MDA, GPx, CAT and TAC) both in plasma and testicular tissue, sperm parameters (motility, vitality, concentration and morphology) and histopathological parameters (MSTD, GECT, Johnson's score, Cosentino's score and testicular cell thickness) were assessed in all groups. Ischaemia/reperfusion significantly increased MDA and decreased GPx, CAT and TAC levels (p < .05). Pretreatment with SMV significantly increased GPx, CAT and TAC levels (p < .05). SMV group increased progressive sperm motility and vitality and reduced non-progressive motility of spermatozoon (p < .05). Testicular torsion significantly decreased all histopathological parameters compared to the sham group (p < .05). SMV pretreatment remarkably increased MSTD, GECT and Cosentino's score in comparison with the TD group (p < .05). A combination of Salvia miltiorrhiza with verapamil could reduce damages triggered by testicular torsion detorsion and improve sperm functionality parameters and oxidative stress defence systems.

The effects of verapamil and heparin co-administration on sperm parameters and oxidative stress in prevention of testicular torsion/detorsion damage in rats.

In this research, the impacts of combined administration of verapamil and heparin on testicular torsion damage were examined. In this experimental study, 30 sexually mature male Wistar albino rats were divided into five equal groups haphazardly (n = 6): Group 1 was the sham group. In group 2, a 2-hr testicular torsion was induced, and thereafter, detorsion was done. Rats in group 3 and group 4 experienced an identical surgical procedure like group 2, but verapamil and heparin were administered in 0.3 mg/kg and 800 IU/kg doses respectively, and in group 5, a combination of verapamil and heparin were administered. Intraperitoneal drug injection in all treatment groups was done 30 min before testicular detorsion. Testicular torsion significantly changed sperm parameters, oxidative stress biomarkers and Cosentino's histological score compared to the sham group (p < .05). All treatment groups reduced testicular damage by decreasing oxidative stress and improving sperm parameters, but heparin and co-administration of verapamil and heparin were significantly better than verapamil injection alone. However, heparin injected group was more effective than other treatment groups (p < .05). Overall, an anticoagulant like heparin is more effective than a calcium channel blocker such as verapamil, and it is more likely to reduce testicular torsion injuries.

The first report of peste des petits ruminants (PPR) in camels (Camelus dromedarius) in Iran.

In mid-July 2013, an outbreak of peste des petits ruminants (PPR) was observed in a herd of camels after they were imported from Kuwait to the Khuzestan province in southwest of Iran. The clinical signs of the affected animals included sudden death, fever, oral erosion, and ecthyma like lesions, yellowish diarrhea, pneumonia and respiratory distress, enlargement of lymph node, severe dehydration, dermatitis, ulcerative keratitis, and conjunctivitis. Necropsy findings included keratoconjunctivitis, congestion and consolidation of the lung, paleness of the liver, and enlargement and edema of lymph nodes. Histopathological exam revealed degeneration and acute hyperemia of the lungs, fatty change and necrotic foci in the liver, tubular necrosis in the kidneys, and necrotic dermatitis. We used immunocapture enzyme linked immunosorbent assay (ELISA) to confirm peste des petits ruminants virus (PPRV) and differentiate it from rinderpest virus. Then virus genome was studied by molecular analysis for detecting of strain and substrain of the virus. Immunocapture ELISA of all specimens reacted positively against PPRV antigens. Also, reverse transcription polymerase chain reaction (RT-PCR) results in the lung and lymph nodes of the dead camels consolidated the cause of disease to be PPRV. The present study is the first report of the PPRV outbreak in camels in Iran.

Comparing the effects of intravenous injection and intranasal atomisation of detomidine in sheep.

BACKGROUND: Detomidine is an α-2 agonist sedative drug which reduces the release of norepinephrine in nerves. Administering this drug through intranasal (IN) route could cause direct transmission to the central nervous system. Therefore, IN administration of detomidine would decrease the side effects and the onset of sedation. OBJECTIVES: In this study, IN administration of detomidine in sheep through an atomiser was compared to its IV administration. METHODS: Fifteen mature female sheep with an approximate weight of 49.53 ± 1.72 kg were used. They were randomly divided into three groups: (1) atomising 10 µg/kg (IND10 ); (2) IV 10 µg/kg (IVD) and (3) atomising 30 µg/kg (IND30 ). Following administration, vital signs, electrocardiographic components, sedative score and biochemistry profile were measured after 15, 30 and 60 min, which were compared with the baseline measures. RESULTS: Bradycardia and the percentage of reduction from the baseline value in the respiratory rate were lower in the IND10 group compared to those in the IVD group. There was no significant difference in terms of the temperature and blood oxygen saturation (SpO2 ) among all the groups (p > 0.05). The level of cortisol declined in all the groups, and in the IND30 (60 min), it was significantly different with the baseline value. The level of glucose increased in all the groups compared to the baseline, which was not significant. Insulin concentration was reduced in all the groups, and in the IND30 group, it changed significantly 60 min after the drug administration. Sedation onset time was faster in the IV group. However, sedation scores between the two administration methods were not different, and only a dose-dependent increase was found in the sedation score in the atomisation group. CONCLUSIONS: Our findings revealed that IN atomisation of detomidine triggers similar sedation as its IV administration, which could be used as an alternative method.

Incidence of iodine deficiency and congenital goitre in goats and kids of Darreh Garm region, Khorramabad, Iran.

One of the thyroid disorders of ruminants is goitre, which is triggered by iodine deficiency. This study evaluates goitre in the goats of the Darreh Garm region in the vicinity of the Khorramabad city. Three goats with congenital enlarged thyroid glands were referred to the Veterinary Teaching Hospital of the Lorestan University with signs of arrhythmia, dyspnea and anorexia. Clinical examination, radiographic and sonographic evaluations were performed. Afterward, a comprehensive visual observation was accomplished in the outbreak region and blood samples were taken for thyroid hormones measurement in does and kids. Moreover, soil and forage samples were collected to assess the iodine concentration and soil parameters. Results indicated that the thyroid hormone concentration in the serum of the affected does and kids were significantly lower than healthy and treated animals. Treatment with sodium thyroxine significantly increased the concentration of T3 and T4 hormones. Pasture (5.28 ± 1.57 mg/kg) and soil (11.0 ± 1.49 mg/kg) iodine levels were lower than normal levels in this region. Histopathological slides of the thyroid glands from the dead kids indicated thyroid follicles with different sizes and hyperplasia of the glands. Overall, a 0.5 mg/kg iodine in the diet meal of the goats needs to be considered for prevention of the iodine deficiency.

The effect of natural infection with different Leptospira interrogans serovars on oxidative stress biomarkers and acute-phase responses in horses and cattle.

Leptospirosis is one of the important zoonotic bacterial diseases with a worldwide distribution that is often subclinical. We aimed to investigate the oxidant/antioxidant balance and acute-phase response in naturally infected horses and cattle with Leptospira interrogans. A total of 600 serum samples from horses and cattle were examined for L. interrogans antibodies using the microscopic agglutination test to determine anti-Leptospira IgG antibodies against a panel of eight important Leptospira antigens in Iran. Then, serum total antioxidant capacity, catalase, glutathione peroxidase, and malondialdehyde activities, and nitric oxide, total protein, serum amyloid A, haptoglobin, and albumin concentrations were measured in seropositive and seronegative samples. Serum catalase activities and malondialdehyde, serum amyloid A, and haptoglobin |concentrations in seropositive cattle and horses were significantly higher (P < .05) than in those that were seronegative. Antibody titers increased from 1:100 to ≥ 1:200 in cattle with L. interrogans infection, resulting in a decrease in the serum total antioxidant capacity (P < .05), an increase in serum glutathione peroxidase (P < .01) activity and nitric oxide (P < .05) , serum amyloid A (P < .01), and haptoglobin (P < .05) concentrations. Following the increase in the agglutinating antibody titers in horses infected with L. interrogans, the serum total antioxidant capacity (P < .01) decreased, and serum nitric oxide (P < .05), malondialdehyde (P < .05), and serum amyloid A (P < .01) concentrations were increased. In this study, horses and cattle had extensive changes in oxidant/antioxidant equilibrium and acute-phase protein concentrations when infected with L. interrogans. We also demonstrated a direct link between antibody titers and the type of leptospiral serovar using serum oxidative and inflammatory markers.

Study on the effect of medicinal leech therapy (Hirudo medicinalis) on full-thickness excisional wound healing in the animal model.

The possible impacts of alternative and conventional medicines on wound healing are now of growing interest. This study aimed to evaluate and elucidate the wound healing activity of medicinal leech therapy in wound excision of the rat model. After a round, full-thickness excision was made in the dorsal region of the body, the animals (n = 30) were randomly divided into three equal groups: I) the treatment group (MLT), where the wounds received leech treatment; II) the positive control group (PC), where the wounds received 1% sodium phenytoin treatment; and III) the negative control group (NC), where the wounds did not receive any treatment. On days 6 and 16, wound biopsy specimens were taken, and prepared sections were stained using various methods. The contraction rate differed significantly (P < 0.05) between the NC group and the other groups. The histopathological evaluation revealed that MLT group showed an accelerated healing process and lower inflammatory response compared to other groups. In ML-treated group maturation and remodeling of collagen had occurred, while in 1% sodium phenytoin treated group, proliferation was the prominent feature. Results showed that the fibroblast was significantly lower in the NC group in comparison to other groups. The number of MNC, s, and PMN, s was significantly higher in the NC group compared to other groups (P < 0.0001). In our study, medicinal leech therapy had a higher success rate in healing for the treatment of excisional wounds in animal models.

Investigating the sperm parameters, oxidative stress and histopathological effects of salvia miltiorrhiza hydroalcoholic extract in the prevention of testicular ischemia reperfusion damage in rats.

AIMS: One of the most common urologic emergencies is spermatic cord torsion, which can damage testicular tissue and reduce fertility. Salvia miltiorrhiza (SM) hydroalcoholic extract possess high antioxidant properties, and its efficacy in ischemia-reperfusion (I/R) injury prevention has been demonstrated in cardiac, renal, and liver tissues. Therefore, the purpose of this study was to assess the protective mechanism of SM extract on testicular I/R damage. MAIN METHODS: 18 mature male Wistar albino rats were randomly divided into 3 groups; with six rats in each group: Group 1 (Sham) was sham-operated. Group 2 (T-D): torsion was performed, and after 2 hours (h) detorsion was done. Group 3 (SM): (200 mg kg-1) SM was intraperitoneally injected thirty minutes before detorsion. Then testicular and epididymal weight and size alterations, sperm parameters (motility, livability, concentration, and morphology), both plasma and testicular tissue levels of malondialdehyde (MDA), catalase (CAT), glutathione peroxidase (GPX), and total antioxidant capacity (TAC) were evaluated. Also, histopathological changes included mean seminiferous tubular diameter (MSTD), testicular capsule thickness (TCT), mean testicular biopsy scoring (MTBS), and germinal epithelial cell thickness (GECT) were examined. RESULTS: Testicular I/R significantly reduced sperm motility, viability, and normality, while SM extract administration remarkably increased sperm motility, and normality (P < 0.05). Induction of testicular T-D caused a significant increment in the level of MDA and notable decline in the levels of GPX, CAT, and TAC both in plasma and testis tissue, whereas administration of SM extract significantly decreased MDA level and increased GPX, CAT, and TAC levels in plasma and testicular tissue (P < 0.05). Histopathological parameters including MSTD, GECT, MTBS, and TCT were significantly lower in the T-D group, while pretreatment with SM extract remarkably increased MSTD, GECT, and MTBS amounts (P < 0.05). CONCLUSION: Since the SM extract increased the activity of antioxidant enzymes, improved sperm parameters and reduced the damage to testicular tissue, therefore, its use as a potent antioxidant in reducing testicular I/R damage is suggested.

Brodifacoum toxicosis and abortion in an Arabian mare.

A 3-year-old pregnant Arabian mare was referred to the Veterinary Teaching Hospital of Shahid Chamran University of Ahvaz with a history of bleeding and rodenticide ingestion. The results of paraclinical examinations showed severe normocytic and normochromic anemia, decreased serum total protein, albumin, and fibrinogen concentrations, increased serum total bilirubin, urea, and creatinine concentrations, as well as increased serum aspartate aminotransferase and creatine kinase activity. Three days after treatment, all the clinical signs were resolved, however, fetus abortion occurred. In order to confirm the suspected cause of abortion and toxicosis, high-performance liquid chromatography was performed on serum sample of mare and liver tissue of the aborted fetus and toxicosis was confirmed. Poisoning with brodifacoum is considered as an important and lethal poisoning for both human being and animals. To our knowledge, this is the first report of spontaneous toxicosis and abortion with brodifacoum. Brodifacoum toxicosis can be effectively managed with early diagnosis, good paraclinical examinations and appropriate treatment.

Evaluation of 5 methods for diagnosing failure of passive transfer in 160 Holstein calves.

BACKGROUND: Inadequate absorption of colostral IgG1 is termed failure of transfer of passive immunity (FTPI). Dairy calves with FTPI have increased mortality and morbidity in their first 6 months of life. OBJECTIVES: This study compared the clinical performance of 5 methods for diagnosing FTPI in Holstein calves. METHODS: An observational study was performed using 160 Holstein heifer calves. Serum was harvested at 48 hours of age, and FTPI was assessed using a digital Brix refractometer for total solids measurements, and digital refractometry and the biuret method to measure serum total protein (STP) concentrations. Serum gamma-glutamyltransferase activity was measured with an automated analyzer, and serum IgG was measured with the zinc sulfate turbidity test and an enzyme-linked immunosorbent assay. Diagnostic test performance was compared with that of the reference method (FTPI defined as a serum total IgG concentration <10 g/L). Test performance was evaluated using the area under the receiver operating characteristic curve, the sensitivity, the specificity, and the positive likelihood ratio at the optimal test cut point, and by calculating the kappa coefficient. RESULTS: A serum digital Brix percentage of <7.8% and an STP concentration of <52 g/L measured using digital refractometry were the best methods to identify calves with FTPI. The STP concentration measured with digital refractometry was 0.1 g/L lower than that measured with the biuret method. CONCLUSIONS: The digital Brix refractometer and the digital refractometer provide accurate and clinically useful methods for identifying dairy calves with FTPI. In this study, the excellent performance of the Brix refractometer was likely due to the use of a fixed sample volume (200 µL) and a uniform sample temperature at the time of measurement.

Vertical transmission of Theileria lestoquardi in sheep.

This is the first report of an outbreak of Theileria lestoquardi abortion and stillbirth in a mob of 450 ewes in July 2012, during which, approximately 35 late-term ewes lost their fetuses over a 5-day period. A dead ewe and her aborted fetus were transported to the Ahvaz Veterinary Hospital for the diagnostic evaluation. The microbial cultures from the ewe vaginal discharges and fetal abomasal contents and the liver were negative. The blood films of the ewe and her fetus contained Theileria piroplasms and the impression smears from ewe liver and fetal spleen were positive for Theileria Koch blue bodies. The DNA was extracted from the liver and spleen of ewe and her fetus, respectively, and analyzed by polymerase chain reaction (PCR) using specific primers derived from the nucleotide sequences of 18S ribosomal DNA (rDNA) gene of T. lestoquardi. A single fragment of 428-bp fragment was amplified. The PCR product was directly sequenced and the alignment of the sequence with similar sequences in GenBank(®) showed 100% identities with 18S rDNA gene of T. lestoquardi. The present study is the first report of the T. lestoquardi vertical transmission that could be related to the abortion.