Evaluation of milk haptoglobin and amyloid A in high producing dairy cattle with clinical and subclinical mastitis in Shiraz

Mastitis is one of the most common diseases in dairy cattle and results in considerable loss of animals. This study was designed to evaluate milk haptoglobin (Hp) and milk amyloid A (MAA) as an inflammatory indicator for clinical and subclinical mastitis of cattle in dairy farms in Shiraz, Iran. Forty-three subclinical mastitic cows with a positive California Mastitis Test (CMT) and no clinical signs of mastitis, 28 clinical mastitic cows, and 10 healthy cows with negative CMT were selected. After confirmation of clinical and subclinical mastitis by bacterial identification, milk samples were taken from four quarters of each cow and mixed, and one sample was taken from the pooled milk. The most dominant isolated bacterium from clinical and subclinical samples was Staphylococcus aureus (n = 25; 35.2%). The most dominant isolated bacterium from clinical (19/28) and subclinical (11/43) samples was Staphylococcus spp. Of isolated bacteria of milk in cattle with clinical mastitis, 67.8% (n = 19) was S. aureus. There was no bacterial growth in 37.1% (n = 16) of cattle with subclinical mastitis. Of isolated bacteria of milk in cattle with subclinical mastitis, 13.9% (n = 6) and 11.6% (n = 5) was S. aureus and Staphylococcus epidermidis, respectively. There were significant differences (P < 0.05) in concentrations of milk Hp, MAA, and somatic cell count between clinically healthy cattle and cows with clinical and subclinical mastitis. The concentrations of milk Hp, MAA, and somatic cell count in clinical mastitic cows were significantly higher than those in subclinical mastitic cows and control group. The optimal cutoff point was set, using the receiver operating characteristic curve analysis method, to >13.43 μg/ml for MAA, >9.71 ng/ml for milk Hp, and >14 × 10⁴ cell per millilitre for somatic cell count with corresponding 100% sensitivity and 100% specificity for MAA, 83.72% sensitivity and 100% specificity for milk Hp, and 88.37% sensitivity and 100% specificity for somatic cell count. The results of this study reveal that MAA is a sensitive factor for diagnosis of subclinical mastitis in cattle.     

Effect of different levels of cigarette smoking on lipid peroxidation, glutathione enzymes and paraoxonase 1 activity in healthy people

Abstract The purpose of this study was to examine the effect of different levels of cigarette smoking on lipid peroxidation, glutathione enzymes and paraoxonase 1 (PON1) activity in a healthy population. The study included 130 subjects who were classified as mild (≤10 cigarettes daily, Group I, n=30), moderate (11–20 cigarettes daily, Group II, n=35), heavy (>20 cigarettes daily, Group III, n=33) and never smokers (controls, Group IV, n=32). Malondialdehyde (MDA) levels, PON1 and erythrocyte glutathione enzyme activities were measured. MDA levels were significantly higher in smokers than never smokers (P<0.05 for Group I, P<0.001 for Group II and III). PON1 activity was significantly lower in heavy smokers (P<0.001). Glutathione peroxidase (GSH-Px) activity was significantly lower in the smokers (P<0.0001). Glutathione reductase (GR) activity was significantly higher in smokers (P<0.0001). MDA levels negatively correlated with PON1 and GSH-PX activities (P<0.01), whereas they positively correlated with GR activities (P<0.001). At every level, cigarette smoking is associated with increased lipid peroxidation and causes an impairment in antioxidant systems.     

L-arginine reverses indomethacin-induced inhibition of inflammatory fluid secretion of the feline gall bladder

Impaired antioxidant defences may predispose to the increased resting and exercise-induced oxidative stress found in patients with insulin-dependent diabetes mellitus (IDDM). We investigated major erythrocyte antioxidant enzyme activities at rest and in response to sustained, moderate intensity physical exercise in young diabetic men (n = 9) previously reported to have markedly elevated plasma lipid peroxidation and blood glutathione levels compared with control men (n = 13) (Laaksonen et al. 1996). At rest, erythrocyte glutathione reductase activity was 15% higher in the diabetic group (P = 0.049). Se-glutathione peroxidase and glutathione-S-transferase activities were similar in both groups. Red cell Cu, Zn-superoxide dismutase and catalase activities were lower in the IDDM group (P = 0.033 and P = 0.023, respectively). After 40 min of exercise at 60% of the subjects' peak oxygen consumption, Se-glutathione peroxidase activity rose by about 14% in the control group (P = 0.003), but not in the IDDM group (P = 0.47). Exercise did not cause significant changes in other enzyme activities in either group. To conclude, lower erythrocyte Cu, Zn-superoxide dismutase and catalase activity in young men with IDDM at rest may contribute to increased oxidative stress. On the other hand, increased glutathione reductase activity may represent a compensatory upregulation of glutathione homeostasis in response to increased oxidative stress. Upregulation of Se-glutathione peroxidase activity in response to physical activity appeared to be impaired in men with IDDM.     

Elevated Serum Cyclophilin Levels in Patients with Severe Sepsis

Several cytokines are considered to be important mediators in the pathophysiology of sepsis. Cyclophilins (Cyps), the main binding proteins for the immunosuppressive drug cyclosporine A, have been suggested to function as cytokines. This study was conducted to determine (i) if serum Cyp levels were elevated in critically ill patients suffering from either sepsis or other life-threatening diseases and (ii) if so, whether there was an association between Cyp levels and a certain diagnosis and/or outcome. Serum samples of 45 patients (22 severe sepsis, 23 other diagnoses) and 17 healthy controls were prospectively analyzed by an enzymatic assay using the ability of cyclophilins to catalyze cis/trans isomerisation of peptidyl-prolyl-peptide bonds (PPIase activity). In addition, western blotting was applied to differentiate both isoforms. PPIase activity was significantly higher in patients with severe sepsis than in patients with other diagnoses (P = 0.004) or in healthy subjects (P = 0.001). There was no difference between healthy subjects and other critically ill patients (P = 0.067). Elevated PPIase activity was associated with high mortality (P = 0.03). It is concluded that Cyps might play a role, probably as mediators in the pathophysiology of sepsis or as symptoms of diagnostic value.     

Incidence Rates of Clinical Mastitis among Canadian Holsteins Classified as High,Average, or Low Immune Responders

The objective of this study was to compare the incidence rate of clinical mastitis (IRCM) between cows classified as high, average, or low for antibody-mediated immune responses (AMIR) and cell-mediated immune responses (CMIR). In collaboration with the Canadian Bovine Mastitis Research Network, 458 lactating Holsteins from 41 herds were immunized with a type 1 and a type 2 test antigen to stimulate adaptive immune responses. A delayed-type hypersensitivity test to the type 1 test antigen was used as an indicator of CMIR, and serum antibody of the IgG1 isotype to the type 2 test antigen was used for AMIR determination. By using estimated breeding values for these traits, cows were classified as high, average, or low responders. The IRCM was calculated as the number of cases of mastitis experienced over the total time at risk throughout the 2-year study period. High-AMIR cows had an IRCM of 17.1 cases per 100 cow-years, which was significantly lower than average and low responders, with 27.9 and 30.7 cases per 100 cow-years, respectively. Low-AMIR cows tended to have the most severe mastitis. No differences in the IRCM were noted when cows were classified based on CMIR, likely due to the extracellular nature of mastitis-causing pathogens. The results of this study demonstrate the desirability of breeding dairy cattle for enhanced immune responses to decrease the incidence and severity of mastitis in the Canadian dairy industry.     

Binding of bovine fibronectin to mastitis-causing Streptococcus agalactiae induces adherence to solid substrate but not phagocytosis by polymorphonuclear cells

Streptococcus agalactiae is frequently associated with mastitis in cattle, and fibronectin is a host protein which interferes with infection, particularly at mucosal sites. The binding of bovine fibronectin to S. agalactiae was investigated by enzyme-linked immunosorbent assay (ELISA). Binding was time-dependent and dose-dependent. Most of the strains tested showed low ELISA activity, but some strains (six out of 17) had much higher activity. The highest-binding strains of S. agalactiae were able to adhere to bovine fibronectin-coated polystyrene. Adherence was inhibited by arginine or gelatin, but was slightly favored by soluble fibronectin. Fibronectin did not contribute to the opsonization (deposition of complement C3) and phagocytosis of the strain which bound the highest amount of fibronectin, even when polymorphonuclear cells were activated with serum-derived chemotaxin (C5a). These data suggest that fibronectin might promote tissue adherence but not phagocytosis of S. agalactiae in the mastitis process.     

Obesity predisposes to the risk of higher mortality in young COVID-19 patients

Although emerging data demonstrated mortality of young COVID-19 patients, no data have reported the risk factors of mortality for these young patients, and whether obesity is a risk for young COVID-19 patients remains unknown. We conducted a retrospective study including 13 young patients who died of COVID-19 and 40 matched survivors. Logistic regression was employed to characterize the risk factors of mortality in young obese COVID-19 patients. Most of the young deceased COVID-19 patients were mild cases at the time of admission, but the disease progressed rapidly featured by a higher severity of patchy shadows (100.00% vs 48.70%; P = .006), pleural thickening (61.50% vs 12.80%; P = .012), and mild pericardial effusion (76.90% vs 0.00%; P < .001). Most importantly, the deceased patients manifested higher body mass index (odds ratio [OR] = 1.354; 95% confidence interval [CI] = 1.075-1.704; P = .010), inflammation-related index C-reactive protein (OR = 1.014; 95% CI = 1.003-1.025; P = .014), cardiac injury biomarker hs-cTnI (OR = 1.420; 95% CI = 1.112-1.814; P = .005), and increased coagulation activity biomarker D-dimer (OR = 418.7; P = .047), as compared with that of survivors. Our data support that obesity could be a risk factor associated with high mortality in young COVID-19 patients, whereas aggravated inflammatory response, enhanced cardiac injury, and increased coagulation activity are likely to be the mechanisms contributing to the high mortality.     

Serum protein electrophoretic pattern in clinically healthy calves and cows determined by agarose gel electrophoresis

The aim of the present study was to determine the physiologic electrophoretic pattern of serum proteins in clinically healthy calves and cows using agarose gel electrophoresis, and to describe the possible influence of age on the concentrations of serum protein fractions in young and adult cattle. Into the evaluation we included 14 clinically healthy calves of a Slovak spotted breed and its crossbreeds at the age of 4–6 months, and 13 clinically healthy cows of the same breed at the age of 3–5 years. Blood serum was analyzed for total serum protein concentrations, and for the absolute and relative values of serum protein fractions using agarose gel electrophoresis. In cows we found significantly higher total serum protein concentrations than in calves (P?<?0.001). Serum protein electrophoresis identified in cows, as well as in calves, six fractions comprising albumin, α₁ and α₂, β₁ and β₂, and γ-globulins. The concentrations of serum protein fractions in cows were 34.8 g/l for albumin, 9.3 g/l for α₁-globulins, 4.9 g/l for α₂-globulins, 6.2 g/l for β₁-globulins, 6.8 g/l for β₂-globulins, and 21.1 g/l for γ-globulins. In calves we observed a marked shift in the concentrations of some protein fractions, with nonsignificantly higher concentrations of α₁-globulins. On the other hand, the values of α₂- and γ-globulins in young animals were significantly lower than those measured in adult ones (P?<?0.01 and P?<?0.001, respectively). Presented results suggest a more marked influence of age on the concentrations of several serum protein fractions in cattle. The relative concentrations of the most of protein fractions showed significant differences between young and adult cattle. Seeing that the analysis of serum proteins and their electrophoretic separation in cattle is less well documented, the study provides important findings for clinicians when evaluating dysproteinemias.     

Associations of long‐chain polyunsaturated fatty acid concentrations with birth outcome in term Indian mothers and their neonates

The purpose of this study was to examine the maternal and cord long‐chain polyunsaturated fatty acid (LCPUFA) concentrations and their associations with birth outcome in term deliveries. Pregnant women (n = 253) delivering at term were divided into two groups based on their babies' birth weights (1) normal birth weight (NBW), that is, ≥2.5 kg (n = 190) and (2) low‐birth weight (LBW), that is, <2.5 kg (n = 63). Each group is further divided into two groups according to the baby's sex, that is, male NBW (M‐NBW), female NBW (F‐NBW), male LBW (M‐LBW), and female LBW (F‐LBW) groups. Maternal plasma docosahexaenoic acid and erythrocyte arachidonic acid (AA) levels were lower (P < 0.05 for both), while cord plasma docosahexaenoic acid and erythrocyte AA levels were higher (P < 0.05 for both) in LBW group when compared to the NBW group. Maternal erythrocyte AA levels were positively associated with birth weight (P = 0.001), while cord plasma docosahexaenoic acid levels were negatively (P = 0.05) associated with birth weight. Reduced maternal and increased cord LCPUFA levels exist in mothers delivering LBW babies, especially in mothers delivering female babies, indicative of sex‐specific effects. Am. J. Hum. Biol., 2011. © 2010 Wiley‐Liss, Inc.     

Effect of different levels of cigarette smoking on lipid peroxidation, glutathione enzymes and paraoxonase 1 activity in healthy people

The purpose of this study was to examine the effect of different levels of cigarette smoking on lipid peroxidation, glutathione enzymes and paraoxonase 1 (PON1) activity in a healthy population. The study included 130 subjects who were classified as mild (20 cigarettes daily, Group III, n=33) and never smokers (controls, Group IV, n=32). Malondialdehyde (MDA) levels, PON1 and erythrocyte glutathione enzyme activities were measured. MDA levels were significantly higher in smokers than never smokers (P<0.05 for Group I, P<0.001 for Group II and III). PON1 activity was significantly lower in heavy smokers (P<0.001). Glutathione peroxidase (GSH-Px) activity was significantly lower in the smokers (P<0.0001). Glutathione reductase (GR) activity was significantly higher in smokers (P<0.0001). MDA levels negatively correlated with PON1 and GSH-PX activities (P<0.01), whereas they positively correlated with GR activities (P<0.001). At every level, cigarette smoking is associated with increased lipid peroxidation and causes an impairment in antioxidant systems.     

The diagnostic value of determination of positive and negative acute phase proteins in milk from dairy cows with subclinical mastitis

Currently, somatic cell count (SCC) and bacterial culture are considered as the gold standard methods for the diagnosis of bovine subclinical mastitis. However, SCC has a low diagnostic accuracy. Therefore, for identification of infected animals, new biomarkers with high diagnostic accuracy are needed. Acute phase proteins (APPs) are proteins that are increased (positive APPs) or decreased (negative APPs) in response to inflammation. The objective of this study was to determine the diagnostic value of milk APPs for the diagnosis of subclinical mastitis in dairy cows. A total of 90 clinically healthy cows were randomly selected. Of these, 52 cows were considered subclinical mastitic based on a SCC higher than 130?×?1,000?cells/mL of milk and positive bacterial culture results of milk samples obtained from at least one of the quarters. Milk amyloid A (MAA) concentration was measured using a commercial ELISA kit and albumin, α-lactalbumin, β-lactoglobulin, and immunoglobulin (Ig) were measured in the whey samples by the use of cellulose acetate electrophoresis. Diagnostic sensitivity and specificity and cutoff points for each test were determined via receiver-operating characteristics analysis. Significant (P?<?0.001) increases in the mean and median concentration of MAA, albumin, α-lactalbumin, and Ig were found in the milk samples collected from cows with subclinical mastitis. MAA was the most accurate test with a diagnostic sensitivity of 92.3% and specificity of 92.1% at cutoff point of >1.6?mg/L. The results of this study showed that determination of MAA can be used as a reliable method for the diagnosis of bovine subclinical mastitis.     

Serum copper, zinc and iron status of various bovine categories on Holstein dairy cattle farms

The aim of this study was to evaluate the blood serum concentrations of copper (Cu), zinc (Zn) and iron (Fe) in various bovine categories reared on industrial Holstein dairy farms. The study was carried out on 12 industrial Holstein dairy cattle farms in various region of Isfahan province, central Iran. Blood samples were collected from 240 dairy cows, 120 dry cows, 120 heifers and 120 calves. The Cu, Fe, and Zn contents were assessed in samples using atomic absorption spectrophotometer. Dry cows and heifers showed the lower Cu concentrations in comparison with dairy cows and calves (P?<?0.05). Heifers and calves showed the higher Zn concentrations in comparison with dairy cows (P?<?0.05). The serum Fe concentrations were significantly higher in heifers than those in dairy and dry cows (P?<?0.05). Dairy cows showed higher Cu and lower Zn and Fe concentrations in comparison with the other categories (P?<?0.05). Cu deficiency was diagnosed in 68% of the examined dairy cows (67% herds), 82% dry cows (92% herds), 75% heifers (92% herds) and 61% calves (58% herds). Only a small number of cattle were Zn- and Fe-deficient while on all farms, Cu deficiency was diagnosed in at least one of the bovine categories. In conclusion, because of the different physiological states of various bovine categories reared on Holstein dairy farms, mineral supplements (Cu, Zn and Fe) should be provided based on physiological requirements of individual bovine categories and the results of those diet analyses.     

Association of Escherichia coli J5-Specific Serum Antibody Responses with Clinical Mastitis Outcome for J5 Vaccinate and Control Dairy Cattle

Dairy cattle in two commercial Holstein herds were randomly selected to be vaccinated twice with J5, at approximately 60 days and 28 days before the expected calving date, or to be untreated controls. Based on whether milk production changed following clinical mastitis or whether cows were culled or died within 30 days after onset, 51 mastitis cases were classified as severe or mild. J5-specific antibody responses were evaluated by enzyme-linked immunosorbent assay of all 32 severe and 19 mild cases. The amounts of J5-specific immunoglobulin M (IgM), IgG1, and IgG2 antibodies in sera from the 27 J5 vaccinates were compared with those of the 24 controls. At drying off (before J5 vaccination), all cows had similar amounts of J5-specific antibody. Immediately after calving (approximately 28 days after the second vaccination), J5 vaccinates had significantly higher production of J5-specific IgG1 and IgG2 than controls. When cows were tested following clinical mastitis, none of the three antibody classes differed significantly between the controls and the vaccinates. Vaccinates that contracted Escherichia coli mastitis had 75% less milk loss than controls. The cows that contracted clinical mastitis later in lactation, the unvaccinated controls, and those infected with E. coli had more milk loss following mastitis. The hazards of being culled for all reasons and of being culled for mastitis were significantly lower for J5 vaccinates. Vaccination with J5 was associated with protection against milk production loss and culling following clinical mastitis, and it was also significantly associated with changes in J5-specific IgM, IgG1, and IgG2 antibodies in sera of vaccinated cows.Coliform mastitis is an important disease complex of dairy cows that is associated with intramammary infections (IMI) with any of several organisms, including Escherichia coli and Klebsiella, Enterobacter, and Citrobacter spp. (9, 11, 20, 22). Clinical mastitis (CM), including abnormal milk, necessity for treatment, milk loss, and death of cattle, can result from coliform mastitis (6, 10, 11, 20, 24). Vaccines against the E. coli rough mutant O111:B4 (J5) bacteria have been in use for 15 years (3, 7). The commercially available vaccines are often called J5 core antigen vaccines, but the antigen(s) and mechanism(s) of J5 immunization have not been clearly identified (2, 23). Associations between J5 vaccination of cattle, various measures of clinical severity of CM, and J5-specific antibodies have rarely been reported for naturally occurring cases of CM (2, 17, 19, 23). Use of survival (time-to-event) analysis to evaluate the relationships between J5 vaccination, antibody production, and time until culling or death has recently been reported for the first time (23). Time-to-event analysis as a measure of incidence of disease or any other event of interest precisely accounts for the time at risk for each animal for that event (5).Naturally occurring cases of bovine CM were studied among J5 vaccinates and controls on two commercial United States dairy farms. The hypothesis was that vaccinates would have more J5-specific antibodies, longer survival times, and less milk production loss following coliform mastitis. J5-specific antibodies (immunoglobulin M [IgM], IgG1, IgG2, and the ratio of IgG1 to IgG2) were therefore compared among vaccinates and controls, as were the relationships of antibody to etiologic pathogen, milk production loss, clinical severity, and time until culling or death. These relationships were deemed important because of their practical relevance and were based upon results of previous intramammary E. coli challenge trials (15, 25).     

The in vitro efficacy of SunSmile® Fruit & Vegetable Rinse against pathogenic strains of Prototheca algae that cause mastitis in cows

The research concerns algae of the genus Prototheca. They are found in the natural environment and they can cause a disease in animals and humans called protothecosis. The aim of the study was to evaluate the in vitro activity of the fruit and vegetable rinse agent SunSmile^® Fruit & Vegetable Rinse (Sunrider International) against P. zopfii isolates. The materials consisted of ten P. zopfii strains isolated from the milk of cows with mastitis. The following antifungal chemotherapeutic agents were also used in the study for comparison: nystatin, ketoconazole, amphothericin B, miconazole, clotrimazole, econazole, fluconazole, and flucytosine. The tube dilution method were used to evaluate the effect of a fruit and vegetable rinse agent and the disc-diffusion method to evaluate the effect of antifungal chemotherapeutic agents on P. zopfii strains. All tested strains of P. zopfii were susceptible to the action of the SunSmile^® agent. The MMC was in the range of 0.0024–0.0190%. The SunSmile^® Fruit & Vegetable Rinse can be used in prevention of mastitis in cows and in human protothecosis due to its safe, natural composition and efficacy.     

Zinc and selenium status in cows with dermatophytosis

Dermatophytosis (ringworm), an infection of the superficial keratinized structures of the skin and hair, is the most common contagious skin disease in cattle. The infectious diseases caused by dermatophytes are mainly related to the enzymes product by these fungi. Conversely, elements such as zinc and selenium are involved in the regulation of immune responses to infection. There are rare reports about the possible role of zinc and selenium concentration in the pathogenesis of cattle dermatophytosis. Thus, this study was conducted in a humid area of Iran on 35 healthy and 35 infected cows. After diagnosis confirmation by direct microscopic examination and fungi isolation via inoculation on Sabouraud dextrose agar using skin scrap and broken hair samples of infected cows, the zinc and selenium concentration of serum and hair in both groups were determined by potentiometric stripping analyzer and atomic absorption spectrometry, respectively. Results showed that serum concentration of selenium and zinc in cattle with dermatophytosis were significantly lower (P < 0.05) than the healthy ones. Although hair concentration of selenium and zinc in infected cattle were lower than the healthy ones, the differences were not significant (P > 0.05). In conclusion, it seems that zinc and selenium have a determinant role in immune status and the response of animal’s immunity system to dermatophytosis.     

Accordance between V̇O2 max and behavior in Ngisonyoka Turkana

This study examined the relationship between maximal aerobic work capacity (maximal oxygen uptake, V?O₂ max) and two variables (occupational group and history of cattle herding) related to the behavior of the Ngisonyoka Turkana, a group of East African pastoralists maintaining a traditional herding lifestyle. Maximal oxygen uptake was measured for 19 men aged 20–44 years using a Monark bicycle ergometer. In line with previous reports for East Africans, there was little difference among mean V?O₂ max for three Ngisonyoka age groups (20–29, 30–39, 40–49 years), P > 0.05. However, V?O₂ max varied with history of cattle herding [defined as either (1) presently herding cattle, (2) had formerly herded cattle, or (3) had never herded cattle], P < 0.01. Maximal oxygen uptake also varied with occupational group (defined as junior herder, senior herder, herdowner [= former herder], or impoverished herdowner [= resumed herder]), P < 0.05. Thus, culturally defined work history and occupational group are positively associated with V?O₂ max, and may be useful in field studies of the effects of past and present activity levels on aerobic capacity. © 1994 Wiley-Liss, Inc.     

Hematological changes of dairy cows in postpartum period and early pregnancy

In this experiment, 50 postpartum healthy Holstein Frisian dairy cows were selected from a large commercial dairy farm. They had normal parturition and had no postpartum problems. The calving number of cows was one to four, and they were inseminated 60 days after parturition. There were no clinical signs of endometritis when cows were examined during routine visits for monitoring fertility. The criteria for selection were that they had a normal parturition history and had calved 25 to 30 days before. They had no mucopurulent discharge from vulva or abnormality in rectal palpation. The cows were evaluated on days 25 to 30 and days 50 to 60 postparturition. Blood samples were collected on sampling days from the coccygeal vein. Hematological parameters were measured by the routine procedures. The health of all uteruses was confirmed by the cytological evaluation of cervical mucosa smear. Cytological samples were obtained from the discharge of cervical mucus and uterine fluid. The concentration of progesterone in serum was measured by radioimmunoassay. There were significant differences in packed cell volume (PCV) between the 25–30 days postpartum, the 55–60 days postpartum, and the pregnant cows. The hematocrit level in the pregnant cows was significantly higher than that in postpartum cows (P < 0.05). The leukocyte count and hemoglobin concentration in the pregnant cows were significantly higher than that in postpartum cows in 25–30 days after parturition. The erythrocyte count in pregnant cows was significantly higher than that of postpartum cows in 55–60 days after parturition. There were significant differences in PCV and hemoglobin in 25–30 and 55–60 days after parturition with the progesterone level of >1 and <1 ng/mL. Hematocrit and hemoglobin concentration in pregnant cows were, respectively, significantly higher than that in postpartum cows with different levels of progesterone (P < 0.05). There were significant differences in segmented neutrophils in the clinically healthy and affected cows by the subclinical endometritis in 25–30 days after parturition. In cows with subclinical endometritis, the percentage and absolute number of segmented neutrophils were significantly higher than in the clinically healthy cows (P < 0.05). There were no significant differences between cows which get pregnant with one and two to three times artificial insemination (P > 0.05).     

Effect of enrofloxacin treatment on plasma endotoxin during bovine Escherichia coli mastitis

OBJECTIVE AND DESIGN: To investigate the effect of enrofloxacin on endotoxin resorption during bovine Escherichia coli mastitis. ANIMALS: 12 healthy early post partum Holstein cows. TREATMENT: Mastitis was induced by intramammary infusion of 10(4) cfu E. coli P4:032. Six cows were treated twice according to the usual enrofloxacin therapy: 5 mg/kg enrofloxacin 1) intravenously at 10 h and 2) subcutaneously at 30 h after challenge. The other 6 cows served as non-treated controls. METHODS: Blood and milk samples were collected at several time points after challenge. LPS in plasma was quantified using the limulus amoebocyte lysate (LAL) assay. The somatic cell count (SCC) and cfu of milk samples were also analysed. RESULTS: Occasional LPS peaks were detected in the plasma of 2 control cows at 6 h post-challenge and of 1 enrofloxacin-treated cow at 10 h post-challenge (P < 0.01 and P < 0.05, respectively, in comparison with time 0), just before enrofloxacin treatment. After enrofloxacin treatment, no significant LPS amounts were detected in the plasma of treated cows, but neither in the control cows. CONCLUSION: During induced coliform mastitis, LPS resorption in plasma occured only sporadically and within 10 h post-challenge. Whereas enrofloxacin treatment clearly limited bacterial growth in milk, significant effects on LPS resorption could not be detected. This suggests that enrofloxacin treatment of E. coli mastitis is predominantly beneficial by its bactericidal activity and is not associated with enhanced resorption of endotoxins.     

Occurrence of <Emphasis Type="Italic">Cryptosporidium parvum</Emphasis> and <Emphasis Type="Italic">Giardia duodenalis</Emphasis> in healthy adult domestic ruminants

To determine the prevalence and intensity of infection of Cryptosporidium spp. and Giardia duodenalis in healthy adult domestic ruminants, faecal samples were collected from 379 cattle of between 3 and 13 years old, 446 sheep and 116 goats selected at random from 60 dairy farms and 38 and 20 herds, respectively, in Galicia (NW Spain). Cryptosporidium spp. oocysts were detected in 32 cows (8.4%), 24 sheep (5.3%) and in nine goats (7.7%) from, respectively, 48.3% of the farms and 34.2 and 30.0% of the herds. The intensity of infection in cows ranged between 25 and 5,924 oocysts per gram of faeces (OPG), whereas in sheep and goats, the number of oocysts shed ranged from 8–515 OPG and from 17–782 OPG, respectively. Parasitization by Cryptosporidium spp. was significantly higher (P < 0.05) in cows than in sheep and goats. G. duodenalis cysts were identified in 101 cows (26.6%), 86 sheep (19.2%) and 23 goats (19.8%) from, respectively, 96.6% of the farms and 92.1 and 90% of the herds. The number of cysts shed by cows ranged between 15 and 3,042 cyst per gram of faeces (CPG), whereas the intensity of infection in sheep and goats ranged from 16–3010 CPG and from 15–1845 CPG, respectively, and was significantly lower (P < 0.05) than in cows and sheep. The number of Cryptosporidium spp. oocysts isolated from sheep and goats was insufficient for successful polymerase chain reaction analysis. Nevertheless, gene sequence analysis of the hsp70 and 18SrRNA genes of Cryptosporidium revealed the presence of only C. parvum in faecal samples from cows. Genotyping studies of the β-giardin and glutamate dehydrogenase genes of G. duodenalis revealed mainly assemblage E of Giardia in cows, sheep and goat faecal samples. Assemblage B of G. duodenalis was also detected in one sheep sample. These animals should be considered as a possible source of cryptosporidiosis and giardiosis, thereby maintaining the infections on farms and in herds.     

Immunodiagnostic identification of dairy cows infected with Prototheca zopfii at various clinical stages and discrimination between infected and uninfected cows

Protothecosis is a severe form of mastitis in cattle that is caused by colorless algae of the genus Prototheca. So far, no suitable serological test for the identification of infected animals is available for routine diagnosis. In this study an indirect enzyme-linked immunosorbent assay (ELISA) for the identification of infected cows and for discriminating among infected cows at various clinical stages was developed. Immunoglobulin G (IgG) in serum and IgA and IgG1 in whey were used as antibody isotypes. The ELISA was evaluated using serum and whey from animals at different clinical stages of infection. A total of 12 cows with acute clinical manifestation of protothecal mastitis, 22 cows with clinical signs of chronic mastitis, 40 Prototheca zopfii-negative cows, and 18 cows with chronic clinical signs and earlier cultures positive for P. zopfii but with presently negative culturing results were investigated. A sensitivity of 96% and a specificity of 94% were calculated for the ELISA based on IgA levels. Intra-assay and interassay variations were calculated to be 6.08 and 6.32%, respectively. Based on these data, this ELISA was found to be suitable for discrimination between infected and uninfected animals and might therefore be useful for screening affected herds.