Coxiella burnetii and Borrelia spp. in peripheral blood of dromedary camels in Fars, Iran: molecular characterization, hematological parameters, and acute-phase protein alterations.

Background: Dromedary camels (Camelus dromedarius) are raised in extremely strict ecological conditions of deserts. Camels are vulnerable to many zoonotic infections. There are limited data on the occurrence of Q fever and borreliosis in camels, in Iran. Aims: The current study was focused on the occurrence of Coxiella burnetii and Borrelia spp. infection in the blood samples of Iranian camels using molecular assays. Effect of the presence of these infections on various hematological factors and some acute-phase proteins (Hp, a1AGP, SAA) were also investigated. Methods: Blood samples were collected from 113 clinically healthy camels to investigate the presence of the infections using nested PCR. Moreover, the sequence of positive samples was analyzed phylogenetically. Routine haematological tests were performed and the concentrations of acute-phase proteins were measured in serum using enzyme immunoassay. Results: PCR result showed that 6.19% (95% CI: 2.53-12.35%) (7/113) of camels were positive for C. burnetii. In addition, sequencing results of the corresponding gene of the outer membrane protein (com1) revealed two different genotypes of C. burnetii agent in camels from Southern Iran. In the PCR assay, Borrelia spp. DNA was not detected in the samples. No significant difference was observed in hematological parameters or acute-phase proteins between positive and negative Q fever camels except for mean corpuscular hemoglobin (MCH) and red cell distribution width (RDW). Conclusion: Clinically healthy camels might be very important reservoirs of zoonotic pathogens. Q fever is not considered a notifiable disease in camels of Iran, and clinical cases may scarcely be recognized by the healthcare system. Due to a lack of adequate information, additional studies on the molecular epidemiology and clinical pathology aspects of C. burnetii infection in Iran are needed.

Effects of citral on serum antioxidant status and liver genes expressions of paraoxonase 1 and nitric oxide synthase in a rat model of streptozotocin-induced diabetes mellitus.

BACKGROUND: Citral (C10H16O) is the main ingredient of Cymbopogon citratus (lemongrass oil) and can reduce the side effects of oxidative stress. Diabetes caused by insulin deficiency induces oxidative stress in the liver. AIMS: This study aimed to investigate the ameliorative effects of citral on selected oxidative parameters and the gene expression of paraoxonase 1 (PON1) and endothelial nitric oxide synthase (eNOS) in a rat model of streptozotocin (STZ)-induced diabetes mellitus. METHODS: Forty rats were divided into four groups at random: control (C), control citral (CC), and two STZ-induced diabetic groups (diabetic (D) and citral diabetic (CD)). After diabetes confirmation (day 7), gavage treatment with citral (300 mg/kg body weight (BW)) was started in the CD and CC groups and continued for two weeks. RESULTS: On day 21 of the study, following treatment with citral for 14 days, the serum levels of total antioxidant capacity (TAC), and PON1 in the CD group were significantly increased compared to those in the D group (P<0.05). While treatment with citral caused a significant decrease in the Malondialdehyde (MDA), and eNOS in the CD group compared to those of the D group (P<0.001). The expression rate of liver PON1 gene was considerably upregulated in the CD group compared to that in the D group (P<0.001); while the opposite was observed for eNOS gene expression. However, there was no significant difference between the CC and C groups in terms of all examined parameters (P>0.05). CONCLUSION: This study showed positive effects of citral on serum antioxidant status and liver gene expression of PON1 and eNOS in diabetic rats.

Alteration of fatty acid profile of milk in Holstein cows fed Bacillus coagulans as probiotic: a field study.

BACKGROUND: Probiotics may improve milk quality and the general health status of animals. AIMS: The effects of dietary Bacillus coagulans PRM101 on milk components, milk fatty acids (FA), and some health indicators of dairy cows were investigated. METHODS: The probiotic was added to the feed of 12 Holstein cows (2 g/cow: 2 × 1011 CFU/cow) for 63 days compared to a control group fed on the basal ration (n=11). Milk and blood samples were taken on days 0, 21, 42, and 63. RESULTS: The yields of milk and energy corrected milk (ECM; computed from milk weight and its fat and protein content) decreased linearly and similarly (P=0.60) in both groups. The treatment cows, however, showed quadratic increases in the weights of milk (P=0.03) and ECM (P=0.04) at d42 of the study. Energy corrected milk (d42, P<0.05) and crude protein content of milk (d42, P<0.05; d63, P<0.1) were higher in the cows receiving the probiotic. The proportions of heptadecanoic (C17:0; P=0.002) and linoleic (C18:2; P=0.077) acids in milk fat (g/100 g fat) were higher in the treatment cows on d63. Milk total antioxidant capacity (TAC), malondialdehyde (MDA), and similarly, amyloid A (AA) and haptoglobin (Hp) of milk and blood were not affected. Total antioxidant capacity and MDA were negatively correlated in the control group (r=-0.669, P=0.005). Heptadecanoic acid correlated negatively with milk MDA (r=-0.611, P=0.035) and positively (r=0.591, P=0.043) with serum Hp in the treatment cows. CONCLUSION: Dietary B. coagulans PRM101 may improve the proportions of C17:0 and C18:2 FA in milk. Some improvements in milk protein and the health status of the cows may also be anticipated.

The effect of L-tryptophan on the food intake, rectal temperature, and blood metabolic parameters of 7-day-old chicks during feeding, fasting, and acute heat stress.

BACKGROUND: Exposure to a high ambient temperature (HT) can cause heat stress, which has a negative impact on physiological functions. L-tryptophan (L-Trp) as a precursor of serotonergic and kynurenine (Kyn) pathways, has a calmative effect during different stress statuses. AIMS: This study was carried out to determine the influence of intraperitoneal injection of Trp on feeding behavior, rectal temperature, and some blood parameters in the heat stress condition. METHODS: L-tryptophan (25 and 50 mg/kg body weight, BW) was administered intraperitoneally during either HT (39°C) or control temperature (CT; 31°C) for 5 h whilst fed or fasted in 7-day-old chicks. RESULTS: L-tryptophan caused elevation in decreased food intake and significantly decreased rectal temperature during acute heat stress at the dose of 50 mg/kg BW. Rectal temperature reduced in the fasted state at the dose of 50 mg/kg BW, and at the dose of 25 mg/kg BW Trp in the fed state in comparison with the other experimental groups. Reduction of serum glucose, triglyceride, and corticosterone levels was seen during the fed state. L-tryptophan had a significant reducing effect on the serum corticosterone level in the fasted state in comparison with the fed state, and also revealed a significant decline at the dose of 25 mg/kg BW on the elevated serum corticosterone under heat stress. CONCLUSION: Administration of L-tryptophan leads to increase cumulative food intake and decrease rectal temperature during heat stress. Also, L-Trp causes to decline increased serum corticosterone level under heat stress and fasted state. These findings indicated the potential regulator role of Trp to modulate stress response in heat-exposed chicks.

The effects of post-partum drops in body condition on indices of energy metabolism in mid-lactation Holstein cows.

BACKGROUND: Evaluation of energy metabolism indices in mid-lactation Holstein cows is critical to monitor health status. Aims: The objective of this study was to assess the effects of low (≤0.75) vs. high (>0.75) drops in body condition score (BCS) until day 60 post-partum on energy metabolism indices during mid-lactation in Holstein cows. METHODS: Twenty-eight Holstein cows were included in the study from the day of calving to day 120 of lactation. Whole blood samples were taken on 60, 90, and 120 days in milk (DIM). Serum was analyzed for insulin, glucose, non-esterified fatty acids (NEFA) and beta-hydroxybutyrate (BHB) levels. RESULTS: In cows with low BCS drop (LoD group), insulin did not change significantly through days 60 to 120 of lactation, but increased in high drop cows (HiD group) (P<0.001). Glucose concentrations decreased linearly in the LoD cows (P=0.039) and showed a quadratic increase in the HiD group on day 90 (P=0.028). Concentrations of non-esterified fatty acids showed both linear (P=0.04) and quadratic (P=0.002) changes in the HiD group. The HiD cows had significantly higher concentrations of insulin on day 120 (P=0.017) compared to the LoD group. Glucose concentration was lower (P<0.01) in HiD cows on 60 DIM. The concentration of non-esterified fatty acids was higher in HiD cows on day 90 (P<0.01). Surrogate indices of insulin resistance (calculated based on the concentrations of the measured metabolites) were different between the groups on day 90, indicating decreased insulin sensitivity in the HiD cows. CONCLUSION: Greater depletion of body reserves during early lactation may result in some inconsistencies in energy metabolism during mid-lactation periods. Controlling BCS loss during early lactation may help alleviate such alterations possibly through modifying insulin sensitivity of the tissues.

Compromised liver functions during the breeding period of clinically healthy Holstein cows.

BACKGROUND: The sub-clinical carry-over effects of post-partum (PP) conditions and the problems independent of parturition may affect the cows' performance during the breeding period. AIMS: It was hypothesized that some mid-lactation cows may have compromised liver functions and calculating liver activity index (LAI; -1.5 to +1.5) may be helpful in detecting such conditions. METHODS: Plasma lipid and protein profiles, retinol and ceruloplasmin were measured in 37 Holstein cows on days 60, 90, and 120 PP. Liver activity index was calculated using the measures of cholesterol, albumin and retinol. The results were interpreted using some health and performance indices. RESULTS: The mean LAI was 0.00 ± 0.61. Sixteen cows (45.7%) were LAI‾ and 19 cows (54.3%) were LAI+. In LAI+ cows the concentrations of cholesterol and albumin were unchanged, but in LAI‾ cows both of them increased through the study (P<0.05). Greater concentrations of low density lipoproteins (LDL) (P<0.01) and lower concentrations of γ- and total globulins (P<0.05; day 60) were detected in LAI+ cows. Ceruloplasmin was not different between the LAI groups with the highest level on day 90. Cholesterol had the strongest and the most repeated correlations with LAI during the study. The correlation of albumin with LAI faded on day 120 and vitamin A had a relationship only on day 60 PP. Triglycerides (TGs), very low density lipoproteins (VLDL) (day 60) and LDL (days 60 and 90) had positive correlations with LAI. All globulin fractions showed negative correlations with LAI on day 60. Seventy percent of the cows without endometritis (day 30 PP) and 33% of the infected cows were LAI+ during the breeding period. About 79% of the cows with body condition score (BCS) loss ≤0.75 (day 60) and 38% of the cows with BCS loss >0.75 were LAI+ during the breeding period. CONCLUSION: Compromised liver functions may exist in dairy cows during the breeding period and may be detected by calculating LAI. The relationship of LAI during the progressed lactation with herd's performance needs to be investigated.

Antioxidant properties of Aloe vera components: a DFT theoretical evaluation.

Prediction of the antioxidant activity of three Aloe vera components (aloesone, aloe-emodin, and isoeleutheol) was performed based on density functional theory calculations using the B3LYP hybrid functional and the 6-311++ G** basis set. Calculation of highest occupied molecular orbital (HOMO), lowest occupied molecular orbital (LUMO), and Egap revealed that aloe-emodin has the lowest Egap value, indicating good antioxidant activity. Also in terms of electron affinity, softness, electrophilicity, and chemical potential, aloe-emodin is a potent structure with potential high radical scavenging activity. Calculation of the ionisation potential revealed that isoeleutherol likely also possesses a high degree of antiradical scavenging. To study the conjugating system of the radicals, density plots of HOMO, natural bond orbital analyses, and spin density plots were used. According to calculations, the isoeleutherol radical is more delocalised and the most stable radical. Calculated proton affinity values revealed that the most probable antioxidant mechanism is sequential proton loss-electron transfer. Our results were compared with available experimental data. Published experimental data were found to correlate well with our theoretical predictions. These results support the usefulness of theoretical calculations not only for identifying potentially useful structures of studied compounds but also for predicting their relative activity.

A field study on glucose, non-esterified fatty acids, beta-hydroxybutyrate and thyroid hormones in dairy cows during the breeding period in Fars province, Iran.

BACKGROUND: It was hypothesized that under apparently good management practices in dairy farms, some cows may not be metabolically perfect during the breeding period and this may affect their performance. AIMS: This study was conducted to assess probable metabolic drawbacks in mid-lactation dairy cows affecting their performance. METHODS: Thirty-seven clinically healthy Holstein cows were assessed for plasma concentrations of glucose, non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHB), thyroxin (T4), and tri-iodothyronine (T3) on days 60, 90 and 120 of lactation. The relationships of the measured analytes with some performance indices were also studied. RESULTS: Continuous declines in plasma glucose (within reference values; P<0.006), T4 (P<0.001) and T3 (0.003) were found during the study. Non-esterified fatty acids showed relatively high levels through the study with a rise at day 90 (P<0.041). Beta-hydroxybutyrate concentrations did not change significantly (P>0.05) but were higher than those reported by others in mid-lactation cows. By progress in lactation 27% of cows had glucose concentrations <2.5 mmol/L, 62% had NEFA concentrations >0.40 mmol/L and 13.5% had BHB levels above 1200 µmol/L, which are the threshold levels of peri- and post-parturient problems. Milk production had negative correlations with glucose and T4 while the correlation was positive with NEFA. The interval between calving to the 1st heat had positive correlations with BHB concentrations. The interval between calving and the 1st insemination was inversely correlated with glucose and positively correlated with NEFA levels. CONCLUSION: The changes of the studied analytes in mid-lactation cows resembled those that would happen during negative energy balance in early lactation cows and feed restriction in mid-lactation cows and may affect the general health and the performance of the cows.

Progesterone concentration and lambing rate of Karakul ewes treated with prostaglandin and GnRH combined with the ram effect during breeding and non-breeding seasons.

The combination of ram effect with two injections of PGF2α 10-days apart and the same protocol plus an additional injection of GnRH prior to the first injection of PGF2α were examined in Karakul ewes during breeding and non-breeding seasons, respectively. Plasma progesterone (P4) concentrations (to detect the presence of active corpus luteum), twin lambing, litter size and synchronization of lambing were evaluated. In each study 70 ewes (2-4 years old) were divided to a treatment (n = 40) and a control (n = 30) group. During the breeding season, on days -10 and 0 before ram release, the treatment group was injected intramuscularly with PGF2α (D-Cloprostenol; 0.15 mg). During the non-breeding season, on day -15 before ram release the treatment group was injected with GnRH (buserelin; 4.2 µg) intramuscularly followed by two injections of PGF2α on days -10 and 0. In both studies, the rams were released into the ewe flock after the second prostaglandin injection (day 0). Blood samples of ewes were collected on days -10, 0, 20 and 70 of the study in breeding season and on days -15, -10, 0, 20 and 70 during non-breeding season. The treatment group had higher P4 concentrations compared to the control ewes on day 0 in the breeding season (5.80 ± 0.61 vs. 5.0 ± 0.93 ng/mL) and day -10 in the non-breeding season (3.50 ± 0.33 vs. 2.70 ± 0.35 ng/mL) though the differences were not significant (P > 0.05). Based on plasma P4 concentrations (>1 ng/mL) on day 70, in the breeding season all control ewes (100%) and 91.9% of the treatment ewes were detected to have active corpus luteum (P = 0.09). An almost inverse result (90% vs. 97.5%; P = 0.2) was detected in the non-breeding season. The lambing rate was higher (P = 0.03) in the treatment group compared to the control ewes during the non-breeding season (90% vs. 70%), but tended to be lower (P = 0.07) in the breeding season (73% vs. 90%). Twin lambing rate was higher in the treatment group compared to the control ewes in the breeding (40.7% vs. 0.0%; P < 0.05) and non-breeding (22.2% vs. 0.0%; P < 0.05) seasons. The litter size of the control and treated ewes were 1.0 ± 0.0 vs. 1.40 ± 0.10 in the breeding and 1.0 ± 0.0 vs. 1.22 ± 0.10 in the non-breeding season (P < 0.05). No effect was observed regard to synchronization of the treated ewes. In the breeding season two injections of PGF2α ten days apart combined with ram effect, may lower the lambing rate, but may enhance twin pregnancies and litter size in Karakul ewes. In the non-breeding season, however, the GnRH-PGF2α treatment plus ram effect may enhance the lambing rate, twin pregnancies and litter size.

Palmitate-induced insulin resistance is attenuated by Pioglitazone and EGCG through reducing the gluconeogenic key enzymes expression in HepG2 cells.

HYPOTHESIS: Palmitate causes insulin resistance (IR) in insulin target tissue. Pioglitazone (an anti-hyperglycemic agent) and epigallocatechin gallate (EGCG, a dietary supplement) can be used for the treatment of type 2 diabetes. However, their molecular effects on gluconeogenesis remain unclear. OBJECTIVE: Hence, we aimed to investigate the simultaneous effect of these anti-hyperglycemic agents on gluconeogenesis through in vitro experiments. METHODS: HepG2 cells were treated with 0.5 mM palmitate, 10 µM pioglitazone, and 40 µM epigallocatechin gallate (EGCG). Gene expression assay was used to investigate the underlying mechanism. Glucose production assay was applied in culture medium to evaluate the activity of gluconeogenesis pathway. RESULTS: Palmitate induced IR could significantly increase G6Pase and PEPCK gene expressions by 58 and 30%, respectively, compared to the control. EGCG reduced the expression of PEPCK and G6Pase by 53 and 67%, respectively. Pioglitazone reduced the mRNA level of PEPCK and G6Pase by 58 and 62% respectively. Combined treatment of insulin-resistant cells with EGCG and pioglitazone significantly decreased the mRNA level of PEPCK and G6Pase by 73 and 80%, respectively. Treatment with palmitate increased glucose production by 50% in HepG2 cells. When the insulin resistant HepG2 cells were treated alone with EGCG and pioglitazone, the glucose production reduced by 50 and 55%, respectively. The combined treatment with EGCG and pioglitazone resulted in 69% reduction in glucose production compared to the palmitate treated HepG2 cells. CONCLUSIONS: These data suggest the additive inhibitory effect of co-treatment with pioglitazone and EGCG on the gluconeogenesis pathway in palmitate-induced insulin resistance HepG2 cells.

Correlation between some hematological parameters, acute phase proteins and serum immunoglobulins in experimental caprine besnoitiosis.

The present study was undertaken to investigate correlation between some hematological parameters, acute phase proteins and immunoglobulins in the experimentally infected goats with Besnoitia caprae from the time of infection till 360 days post infection (DPI). Six male goats, approximately 12-16 months old, were inoculated subcutaneously with approximately 1.3 × 10(8) bradyzoites of B. caprae and blood samples were collected at weekly intervals from the jugular vein of the goats. Total leukocyte count and differential leukocyte counts were determined. Acute phase proteins (APPs) including serum amyloid A (SAA), haptoglobin (Hp), fibrinogen and ceruloplasmin were undertaken at weekly intervals. We evaluated an enzyme-linked immunosorbent assay (ELISA) (using a somatic antigen of bradyzoite) to detect anti-B. caprae antibodies in caprine sera. Cysts were present in the skin biopsies of the distal parts of the leg of the infected goats from 28 DPI. From 30 to 360 DPI, results showed that the APPs concentrations including SAA, Hp, fibrinogen and ceruloplasmin were enhanced in the serum of infected goats. However, there were some variation in hematological parameters; the differences were not significant with those of the normal values. Some variations were seen in the levels of specific antibodies against this parasite and they had correlation with some hematological parameters and acute-phase proteins.

The effects of probiotic, prebiotic and synbiotic diets containing Bacillus coagulans and inulin on rat intestinal microbiota.

An in vivo experiment was conducted to study the effects of probiotic Bacillus coagulans spores, with and without prebiotic, inulin, on gastrointestinal (GI) microbiota of healthy rats and its potentiality to survive in the GI tract. Forty-eight male Wistar rats were randomly divided into four groups (n=12) and fed as follows: standard diet (control), standard diet supplied with 5% w/w long chain inulin (prebiotic), standard diet with 10(9)/day spores of B. coagulans by orogastric gavage (probiotic), and standard diet with 5% w/w long chain inulin and 10(9) spores/day of B. coagulans by orogastric gavage (synbiotic). Rats were fed the diets for 30 days. At day 10, 20 and 30 of experiment, 24 h post administration, four rats from each group were randomly selected and after faecal collection were sacrificed. Small intestine, cecum, and colon were excised from each rat and used for microbial analysis. Administration of synbiotic and probiotic diets led to a significant (P<0.05) increment in lactic acid bacteria (LAB), total aerobic and total anaerobic population compared the prebiotic and control diets. A significant decrease in Enterobacteriaceae counts of various segments of GI tract (except small intestine) in synbiotic, probiotic and prebiotic fed groups were also seen. The obvious decline in spores count through passing GI tract and high surviving spore counts in faecal samples showed that spores are not a normal resident of GI microbiota and affect intestinal microbiota by temporary proliferation. In conclusion, the present study clearly showed probiotic B. coagulans was efficient in beneficially modulating GI microbiota and considering transitional characteristics of B. coagulans, daily consumption of probiotic products is necessary for any long-term effect.

Udder edema and association with some serum biochemical measurands and dietary factors in first calving cows.

The aims of this study were to determine some major biochemical alterations observed in first calving cows with udder edema during the periparturient period and to detect some associations between dietary factors and the disease. For that, the concentrations of some electrolytes (Na(+), K(+), Cl, Ca(2+), P and Mg(2+)), lipid (triglycerides and cholesterol) markers and lipoproteins (HDL, LDL and VLDL) and total proteins were measured in serum samples collected from 70 first calving cows (35 with udder edema and 35 healthy ones) whereas the percentages of dry matter and crude proteins and the electrolyte (Na(+), K(+), Cl, Ca(2+), Mg(2+), P and sulphates) amounts in feed rations were determined in parallel. The total protein, the calcium and the phosphorus as well as the concentrations of lipid markers and lipoproteins (HDL and LDL) were significantly decreased in first calving cows with udder edema compared to the healthy ones and these biochemical alterations were correlated with a reduced dry matter content and an electrolyte desequilibrium mainly involving Na(+) and Cl(-) in feed rations distributed to the cows with udder edema. To our knowledge the mechanism(s) of physiologic udder edema is uncertain and the obtained results suggest that a transient liver dysfunction (decreased total protein and LDL) probably linked to a feed ration deficient in dry matter may be involved in the aetiology of the udder edema in first calving cows.

Genetic characterization and phylogenetic analysis of Eimeria arloingi in Iranian native kids.

Among the 16 species of Eimeria from goats, Eimeria arloingi and Eimeria ninakohlyakimovae are regarded as the most pathogenic species in the world and cause clinical caprine coccidiosis. E. arloingi is known to be an important cause of coccidiosis in Iranian kids. Molecular analyses of two portions of nuclear ribosomal DNA (internal transcribed spacer1 (ITS1) and 18S rDNA) were used for the genetic characterization of the E. arloingi. Comparison of the sequencing data of E. arloingi obtained in the present study (ITS1: KC507793 and 18S rDNA: KC507792) with other Eimeria species in the GenBank database revealed a particularly close relationship between E. arloingi and Eimeria spp. from the cattle and sheep. The phylogram based on the ITS1 sequences shows that the E. arloingi, Eimeria bovis, and Eimeria zuernii formed a distinct group separate from the other remaining Eimeria spp. in cattle and poultry. In pairwise alignment, 18S rDNA sequence derived from E. arloingi showed 99% similarity to Eimeria ahsata with differences observed at only three nucleotides. This study showed that the ITS1 and 18S rDNA gene are useful genetic markers for the specific identification and differentiation of Eimeria spp. in ruminants.

Dynamics of the enzymatic antioxidants during experimental caprine coccidiosis.

Caprine coccidiosis, caused by coccidian parasites from genus Eimeria, is considered as one of the most common and significant diseases in goats worldwide. The aim of this experiment was to investigate the responses of the enzymatic antioxidant systems during experimental coccidiosis. For this purpose, 20 newborn kids were selected. Ten were infected with sporulated oocysts of the most pathogenic species of Eimeria, and the remainder served as the control. Blood samples were taken at 0 (before inoculation), 3, 7, 14, 21, 28, and 35 days post-infection (dpi), and antioxidant-oxidant-related parameters were measured. Our data showed that the activities of the main erythrocyte antioxidant enzymes revealed significant declines at 7 dpi. These decreases were more evident at 14 to 21 dpi and then gradually enhanced to the normal values until 35 dpi; however, total antioxidant capacity revealed a remarkable decrease at 7 dpi and remained on the same level toward the end of the study. By contrast, serum levels of malondialdehyde (a biomarker of lipid peroxidation) and total homocysteine significantly increased at 21 and 14, 21, and 28 dpi, respectively. These observations suggest that caprine coccidiosis can impair the major antioxidant systems leading to remarkable oxidative damages during the infection. Furthermore, oxidative injuries could have a considerable linkage to the pathogenesis of Eimeria parasites.

Acute phase response in lame cattle with interdigital dermatitis.

This study was undertaken to evaluate acute phase response via assessing the concentration of serum sialic acids (total, lipid-bound and protein-bound), inflammatory mediators (IFN-γ and TNF-α) and acute phase proteins [haptoglobin (Hp) and serum amyloid A (SAA)] in lame cattle with interdigital dermatitis. Fifteen hoof scrapings from lame cows were collected from eight commercial dairy farms. As a consequence of the difficulty in culturing and isolation, a PCR technique was used to detect the organism. None of the colonies on enriched blood agar was identified as Fusobacterium necrophorum. Four (26.6%) out of the 15 hoof scrapings examined tested positive for the presence of the lktA gene (402 bp) of F. necrophorum. It seems that culture cannot be considered as the gold standard method for F. necrophorum isolation. Molecular detection is suggested as an alternative method. In the blood serum of different groups of animals (control, lameness and F. necrophorum-positive lameness) Hp, SAA, total sialic acid, lipid-bound sialic acid, and protein-bound sialic acid, and IFN-γ and TNF-α were measured using validated standard procedures. All parameters were significantly higher in the lameness group and the F. necrophorum-positive lameness group compared with the healthy group (P < 0.01 in all cases). Mean SAA concentrations in the lameness group and the F. necrophorum-positive lameness group was relatively 4.6 and 8.0 times higher than the control group. Corresponding measures for Hp indicate a 3.3 times increase in the lameness group compared to the control. In the lameness group, significant associations were observed for Hp with PBSA, SAA with TSA, TSA with PBSA, TSA with LBSA, PBSA with LBSA, and SAA with IFN-γ.

Zinc, copper, iron, and chromium concentrations in young patients with type 2 diabetes mellitus.

Homeostasis of trace elements can be disrupted by diabetes mellitus. On the other hand, disturbance in trace element status in diabetes mellitus may contribute to the insulin resistance and development of diabetic complications. The aim of present study was to compare the concentration of essential trace elements, zinc, copper, iron, and chromium in serum of patients who have type 2 diabetes mellitus (n = 20) with those of nondiabetic control subjects (n = 20). The serum concentrations of zinc, copper, iron, and chromium were measured by means of an atomic absorption spectrophotometer (Shimadzu AA 670, Kyoto, Japan) after acid digestion. The results of this study showed that the mean values of zinc, copper, and chromium were significantly lower in the serum of patients with diabetes as compared to the control subjects (P < 0.05). Our results show that deficiency of some essential trace elements may play a role in the development of diabetes mellitus.

Acute phase response in cattle infected with Anaplasma marginale.

This study was undertaken to evaluate the acute phase responses via the assessment of the concentration of serum sialic acids (total, lipid bound and protein bound), inflammatory mediators (IFN-γ and TNF-α) and acute phase proteins (Hp and SAA) in 20 adult crossbred cattle naturally infected by Anaplasma marginale. The infected animals were divided into 2 subgroups on the basis of parasitemia rate (<20% and >20%). Also, as a control group, 10 clinically healthy cattle from the same farms were sampled. Our data revealed significant decreases in red blood cell count (RBC), hematocrite (PCV) and hemoglobine (Hb) in infected cattle compared to healthy ones. Conversely, the concentrations of Hp, SAA, ceruloplasmin, fibrinogen, serum sialic acids and the circulatory IFN-γ and TNF-α were increased in the diseased cattle (P<0.05). In addition, it was evident that the progression of parasitemia in infected cattle did not induce any significant alterations in the hematological indices (RBCs, PCV and Hb) and the concentrations of Hp, SAA, ceruloplasmin and fibrinogen. SAA was the most sensitive factor to change in the diseased cattle. Therefore, increase in SAA concentration may be a good indicator of inflammatory process in cattle naturally infected with Anaplasma marginale.

In vitro cultivation of axenic amastigotes and the comparison of antioxidant enzymes at different stages of Leishmania tropica.

The present study aimed to establish a simple method to yield large amounts of Leishmania tropica amastigote-like forms in axenic cultures and to compare the superoxide dismutase (SOD) and glutathione peroxidase (GPX) enzymes at different stages of L. tropica. Different culture conditions were tested to find the optimum condition of axenic amastigotes generation. Superoxide dismutase (SOD) and glutathione peroxidase (GPX) activities were determined at logarithmic and stationary phases and axenic amastigote stage of the parasite. A high proportion (88%) of amastigote-like forms of L. tropica was observed in BHI medium supplemented with 20% FCS, pH 4.5, and incubated at 37ºC with 5% CO(2). The results showed that SOD activity was at the lowest level in the logarithmic phase of promastigotes and increased towards the stationary phase of promastigotes and amastigote stage. The results showed that the optimum condition for differentiation of L. tropica promastigotes to axenic amastigotes was BHI medium containing 20% FCS at pH 4.5, incubated at 37ºC in the presence of 5% CO(2). It seems that SOD, but not GPX is a major determinant of intracellular survival of the parasite.

Alterations of erythrocyte antioxidant mechanisms: antioxidant enzymes, lipid peroxidation and serum trace elements associated with anemia in bovine tropical theileriosis.

In order to investigate the alterations of erythrocyte protective antioxidant mechanisms, lipid peroxidation and trace elements associated with anemia in bovine tropical theileriosis, an infected group comprised of 50 crossbred Holstein cattle, about 1-2 years old, naturally infected with Theileria annulata, were divided into 4 subgroups according to their parasitemia rates (<1%, 1-3%, 3-5%, >5%) and also 10 healthy cattle as control were selected. Blood samples were taken and hematological parameters, the activities of antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase and serum concentrations of some antioxidant trace elements (copper, iron, zinc, manganese and selenium) were measured. As an index of lipid peroxidation, the level of Malondialdehyde (MDA) was also determined. The results showed a conspicuous decrease in the activities of SOD, GPX and catalase (P<0.01), and a significant decrease in the serum concentrations of Cu, Zn, Mn and Se in cattle with higher than 1% parasitemia (P<0.05) compared to the control. In addition, remarkable elevations in the MDA level (P<0.01) and serum concentration of iron (P<0.05) were observed in the infected animals. These findings pointed to the occurrence of exacerbating oxidative injuries to erythrocytes during parasitemia. Furthermore, it can be concluded that infection with T. annulata can interfere with protective antioxidant mechanisms of RBCs against oxidative damages, which promote the development of anemia.