BACKGROUND: Cryptosporidium parvum is a protozoan parasite which belongs to apicomplexa phylum. The parasite infects both wild and domesticated animals and human beings as well. OBJECTIVES: The purpose of the present study was to detect oocyst shedding and diarrhea pattern in experimental cryptosporidiosis and their correlation with weight loss in neonatal calves. METHODS: Twelve Holstein calves of both sexes were obtained at birth from dairy farm and randomly divided into two groups of 6 calves. Six calves were orally infected with 107 C.Parvum oocysts at the 12h post parturition. The control group was not infected. Clinical signs were examined and fecal samples were collected by the rectal examination twice a day. All calves were weighed from day 0 to day 30 with 3 days intervals to determine effects of cryptosporidiosis on weight gain. RESULTS: All infected calves were noticeably depressed and had a decreased appetite from 3 days post inoculation (DPI) while they received colostrum. Subsequently, watery diarrhea with clumps of mucus and yellow or pale changes of feces color were observed. The infected calves have had diarrhea for 5-8 days that remarkably had got dehydrated. The most severity of diarrhea was 4-6 DPI. Oocyst excretion started 4 DPI, peaked at 6 DPI (60.48×106±9.03oocysts/g feces) and continued until 11 DPI. Control calves had no diarrhea and other clinical signs during the whole period of the trial. The mean weight gain of control group was significantly higher than inoculated group during experiment (p<0.001). The Weight of the infected calves was retarded until 9 days old and then risen subsequently. CONCLUSIONS: Present study showed the role of C.Parvum as the primary cause of diarrhea and weight loss among neonatal calves.

BACKGROUND: Diagnosis of Cryptosporidium species based on morphological characteristics is very limited and has no taxonomic value alone, and therefore the use of molecular methods removes these limitations to some extent. OBJECTIVES: The aim of this study was to determine the predominant Cryptosporidium genotypes in calves with diarrhea. METHODS: Study were conducted in calves aged less than 3 months for a period of 2 years. During the study period, 160 dung samples were collected from neonatal calves and examined first microscopically and then by molecular techniques. Stools were analyzed for the presence of Cryptosporidium oocysts by Sheather's Sugar Flotation Solution followed by Ziel-Neelsen staining method. DNA of parasite was extracted and multiplex nested-PCR protocol basis on the 18srRNA were done to identify three cattle-adapted species (C. andersoni, C. bovis and C. ryanae) plus the zoonotic species C. parvum. RESULTS: 110 fecal samples were collected from livestock in Alborz province and 50 fecal samples were collected from livestock in Shahroud city. Of the 160 animals examined, 90 were female and 70 were male. In total, out of 160 animals examined, 85 cases (53.12%) had diarrhea, of which 55 cases (34.37%) were positive using Ziel-Neelsen staining. Since all positive cases were related to diarrhea samples and related to calves under one month old, a significant relationship was observed between diarrhea status and the presence of this parasite (p < /em><0.05). In terms of seasonal distribution, no difference was observed in the rate of diarrhea and positive parasitic cases. The presence of 305 bp band in all Ziel-Neelsen positive samples confirmed the presence of C. parvum in all samples. CONCLUSIONS: Neonatal calves are more likely to be infected with Cryptosporidium parvum, as confirmed by the present study.

BACKGROUND: Heat treatment of colostrum has been suggested as a control measure to eliminate or reduce the transfer of colostrum-borne pathogens to dairy calves.Objectives: The aims of this study were to determine the effects of on-farm heat treatment of bovine colostrum on colostral bacterial counts and IgG concentration and evaluation of passive transfer of immunity in neonatal dairy calves. Methods: Ninety-six L of first milking colostrum was collected from Holstein cows and pooled to create a uniform batch. Twenty-four calves were enrolled in 4 treatment groups before suckling occurred and fed raw colostrum (n=6), heat-treated colostrum at 60 ºC for 30 min (n=6), heat-treated colostrum at 60 ºC for 60 min (n=6) and heat-treated colostrum at 60 ºC for 90 min (n=6). Colostrum samples were collected before and after heat treatment and cultured for total bacterial count and analyzed for total IgG concentration. For the first and second feeding 2 L of colostrum was bottle fed by 2 and 12 h of age respectively. Serum samples were collected from calves at 0 h (precolostrum) and 6, 24, 48, 72 h (postcolostrum) and analyzed for serum total protein and IgG concentrations. Results: Heat treatment of colostrum at 60 ºC for 30 and 60 min reduced total bacterial count, yet maintained colostrul IgG concentration compared to the control. There was no difference between treatment groups when examining serum total protein and IgG concentrations, but apparent efficiency of IgG absorption was significantly greater at 6 h in calves that were fed heat-treated colostrum compared to calves fed raw colostrum. ConclusionS: There was no effect of on-farm batch heat treatment of colostrum at 60 ºC till 90 min on serum concentration of IgG.

BACKGROUND: Annexins (including annexin A1 and annexin A2) are important proteins which have some roles in organisms such as intracellular signal conduction, membrane cellular skeletal connection, cellular proliferation and differentiation, especially inhibitory function in inflammation processing. Pasteurella multocida is the most common bacterial pathogen and has high prevalence in pneumonia. Objectives: This study was aimed to determine experimentally annexin A1 and annexin A2 in the serum of calves affected by Pasteurella multocida pneumonia. Methods: In this research, 10 male calves (2 - 4 months) were allocated to two equal groups, one group as the case: 300 ml in dilution 2×109 CFU Pasteurella multocida bacteria and the other as control group: 300 ml normal saline inoculated by special lavage catheter through oral to trachea. Clinical scores were recorded based on available tables. In treatment group, about 18 to 24 hours after inoculation and synchronous with observation clinical signs changes, bronchoalveolar lavage for cytology and bacteriology examination were done of fluids results from washing. Blood sampling was taken from calves jugular vein in both groups then blood serums were examined by using ELISA kits. Results: The rates of annexin A1 and annexin A2  in blood serum of treatment group showed significant increase (using independent t test) compared to control group (p<0.05). Conclusions: It seems these annexins (annexin A1 and annexin A2) can be used as important biomarkers in blood serum to diagnose inflammation processes such as pneumonia.

Receptors for opsonins, such as complement component C3b (CR1) and immunoglobulins, Fc receptors, interact with adhesion glycoproteins in mediating immune functions. Defects in expression of the adhesion glycoproteins CD11/CD18 results in severely hampered in vitro and in vivo adherence-related functions of leukocytes. Little is known regarding the effect of leukocyte adhesion deficiency (LAD) on ligand binding and receptor expression. We investigated the binding and expression of CR1 and Fc receptors by bovine neutrophils isolated from dairy calves suffering from LAD, compared with clinically normal (hereafter referred to as normal) age-matched calves. Neutrophils were also assayed for endogenously bound IgG and IgM and for exogenous binding of C3b, IgG1, IgG2, IgM, and aggregated IgG (aIgG), using flow cytometry. Luminol-enhanced chemiluminescence (CL) production in response to IgG2 opsonized zymosan was studied, and specific inhibition of CL was used to determine the specificity of IgG2 binding. Activation of protein kinase C with phorbol myristate acetate was used to determine the effect of cellular activation on expression of CR1. A greater percentage of neutrophils from normal calves bound C3b than did neutrophils from LAD-affected calves. Receptor expression was similar. Activation with phorbol myristate acetate resulted in increased expression of CR1 on neutrophils from normal and LAD-affected calves, but expression was almost twofold greater on neutrophils from normal calves. There was no difference between LAD-affected and normal calves in percentage of neutrophils that bound endogenous IgG and IgM. A greater percentage of neutrophils from normal calves bound exogenous IgM than did neutrophils from LAD-affected calves. Receptor expression for aIgG was greater on neutrophils from LAD-affected calves than on those from normal calves.

During growth, central venous, right ventricular, pulmonary arterial, Pulmonary capillary wedge, and systemic arterial pressures, heart rate, and cardiac Output were repeatedly measured in 41 Friesian calves, considered as having conventional muscular conformation, and in 19 Belgian White and Blue double-muscled calves. A total of 123 and 70 recordings were collected in conventional and double-muscled calves, respectively. These circulatory indices were calculated: stroke volume, cardiac and stroke indices, pulmonary and systemic pulse pressures, pulmonary and systemic vascular resistance indices, and right and left ventricular work indices. Results indicated that systemic arterial and pulse pressures, as well as cardiac output, stroke volume, cardiac and stroke indices, and right and left ventricular work indices were significantly (P less than or equal to 0.05 to 0.001) lower but, in contrast, pulmonary and systemic vascular resistance indices were significantly (P less than or equal to 0.001) higher in double-muscled than in conventional calves. Right-sided vascular pressures and heart rate were similar in the 2 groups. These results indicated that global cardiac performance may be considerably poorer in double-muscled calves. Diminished cardiac performance of double-muscled calves appears to be related neither to relative bradycardia nor to reduced ventricular preload. The potential role of increased ventricular afterload or of reduced myocardial contractility in double-muscled cattle should be determined by direct measurements.

A study was conducted to investigate whether aspiration of amniotic fluid is associated with a deleterious effect on absorption of colostral immunoglobulins or on blood gas and acid-base values of healthy newborn calves. Fourteen calves purchased from commercial sources were transported to a research facility immediately after birth and fed colostrum with known concentrations of immunoglobulins. Blood samples for gas analyses were collected within 5 hours of birth, 24 hours later, and prior to euthanasia. Between 3 and 5 days of age, calves were euthanatized by an overdose of barbiturates. Eleven calves had evidence of bronchoaspiration of amniotic fluid, as determined by presence of meconium, squamous epithelium, or keratin in histologic sections of fixed lung or by cytologic analysis of bronchoalveolar lavage fluid. Blood gas tensions and pH were within reference ranges in 11 of 14 calves. Aspiration of amniotic fluid could not be linked to any specific changes in blood gas tensions, acid-base status, or absorption of colostral immunoglobulins. Presence of keratin and meconium in the lungs often was accompanied by mild exudative alveolitis and focal atelectasis. It was concluded that aspiration of small amounts of amniotic fluid with or without meconium is common in calves and is not associated with hypoxemia, respiratory acidosis, or failure of passive transfer.

The metabolic responses of equine articular cartilage to incubation with bacterial lipopolysaccharide (LPS) were studied, using explant cultures of articular cartilage obtained from the metatarsophalangeal joints of 15 horses, age of which ranged from 3 months to 20 years. For comparison, explants were also established from the metatarsophalangeal joints of 3 calves. Explants were cultured for 3 days in medium containing various concentrations of LPS from 0 (control) to 100 microgram/ml. Glycosaminoglycan (GAG) released during the 3-day incubation was determined by a spectrophotometric assay, using the dye 1,9-dimethylmethylene blue. Newly synthesized GAG content was assayed by measuring [35S]sulfate incorporation during a 3-hour pulse labeling period. In addition, prostaglandin E2 (PGE2) synthesis was quantified, using a [3H]PGE2 radioimmunoassay kit and magnetic separation. Finally, explants from 3 animals were used to evaluate the effect of supplementing culture medium with 5% serum on the response of explants to LPS, and explants from 1 horse were used to compare responses to stimulation with LPS derived from 2 bacterial sources. Equine explants cultured with bacterial LPS had a dose-dependent decrease in synthesis and increase in release of GAG, and these responses were significantly (P < 0.0001) greater in explants from younger horses. In addition, equine explants had a significant (P = 0.0001) dose-dependent increase in concentration of PGE2 released into the culture medium in response to incubation with LPS. Comparison of data for GAG synthesis from equine and bovine explants revealed a significant (P = 0.025) difference in responsiveness to LPS between the 2 species. Equine explants tended to have a greater suppression of GAG synthesis in response to incubation with increasing concentrations of LPS than did age-corrected bovine samples.

Two hundred ninety-six Eschericbia coli isolates from feces or intestines of calves with diarrhea were hybridized with 7 gene probes. One probe (the eae probe) was derived from the eae gene coding for a protein involved in the effacement of the enterocyte microvilli by the group of bacteria called attaching and effacing E coli (AEEC), and 2 probes were derived from genes coding for the Shiga-like toxins (SLT) 1 and 2 produced by the verocytotoxic E coli (VTEC). The other 4 probes were derived from DNA sequences associated with the adhesive properties of enteroadherent E coli (EAEC) to cultured cells (the EAF probe for the localized adherence pattern, probes F1845 and AIDA-1 for the diffuse adherence pattern, and the Agg probe for the aggregative adherence pattern). Hybridization results for the eae probe were in agreement, for all but 1 of the 8 isolates, with previously published phenotypic results of microvilli effacement. The latter was previously reported as effacing the microvilli of calf enterocytes, but was eae probe-negative. Two classes of isolates hybridized with the eae probe. Members of a first class (60 isolates) additionally produced a positive signal with 1 or both of the SLT probes (VTEC-AEEC isolates). Isolates hy- bridizing with the eae and the SLT1 probes were the most frequent: 56 isolates (ie, 93% of all VTEC-AEEC). Members of the second class (10 isolates) failed to hybridize with either SLT probe (non-VTEC-AEEC isolates). Most isolates of these 2 classes belong to only 4 serogroups: O5, O26, O111, and O118. In addition to these 2 AEEC classes, a VTEC class (20 isolates) was observed. Such isolates were positive with 1 or both SLT probes, but were negative with the eae probe. All but 1 isolate belonged to serogroups not found among the AEEC isolates. Only 7 of all AEEC and VTEC isolates were positive with the EAF, the F1845, or the AIDA-1 probe, and none were positive with the Agg probe. On the other hand, 32 non-VTEC, non-AEEC isolates were po.