Serum periostin levels in early in pregnancy are significantly altered in women with miscarriage.

BACKGROUND: Miscarriage is a common complication in pregnancy and there is still a lack of biomarkers usable in asymptomatic patients before the event occurs. Periostin (PER), whose levels rise particularly during injury or inflammation, has been shown to play an important local role in implantation and early embryonic development. As PER has been described as a biomarker in various medical conditions we intended to evaluate if changes in PER serum levels may help to identify women at risk for spontaneous abortion in the first trimester. METHODS: Women between 18 and 42 years without confounding comorbidities who conceived by IVF/ICSI and ovarian hyperstimulation were analysed in the study after informed consent. Maternal serum samples from 41 patients were assessed at the time of pregnancy testing (PT) and the following first ultrasound checkup (US). Patients were subsequently divided in two groups: (1) patients with subsequent miscarriage in the first trimester (n = 18) and (2) patients with ongoing pregnancy (n = 23), allowing for statistical analysis and investigating the change of PER levels per individual. PER levels were measured using enzyme-linked immunosorbent assay. Statistical analysis was performed using the Fisher exact and Student's t test. p ≤ 0.05 was considered to be significant. RESULTS: There was no significant difference concerning possible confounders between the two groups. We did not find any significant difference in PER levels at the time point of PT or US. By investigating the interindividual changes of PER between the two time points however, we observed that patients with a following miscarriage showed increasing levels of PER at the time point of PT compared to US in contrast to patients with an ongoing pregnancy who demonstrated a decrease in PER levels. These alterations were significant in the absolute as well as in the relative comparison. CONCLUSION: The relative expression of PER between PT and US is significantly altered in asymptomatic women with subsequent miscarriage compared to women with ongoing pregnancy. Therefore systemic PER levels might represent a potential promising biomarker for the assessment of pregnancy outcome. TRIAL REGISTRATION: Not applicable.

Effect of dietary energy and protein content on growth and carcass traits of Pekin ducks.

A study was conducted to determine the influence of dietary energy and protein concentrations on growth performance and carcass traits of Pekin ducks from 15 to 35 d of age. In experiment 1, 14-d-old ducks were randomly assigned to 3 dietary metabolizable energy (11.8, 12.8, and 13.8 MJ/kg) and 3 crude protein concentrations (15, 17, and 19%) in a 3×3 factorial arrangement (6 replicate pens; 66 ducks/pen). Carcass characteristics were evaluated on d 28, 32, and 35. In Experiment 2, 15-d-old ducks (6 replicate cages; 6 ducks/cage) were randomly allotted to the 9 diets that were remixed with 0.5% chromic oxide. Excreta were collected from d 17 to 19, and ileal digesta was collected on d 19 to determine AMEn and amino acid digestibility. In Experiment 1, there were interactions (P<0.05) between dietary metabolizable energy and crude protein (CP) on body weight (BW) gain and feed intake, wherein BW gain increased more to increasing dietary CP as dietary metabolizable energy increased. However, feed intake was only influenced by dietary crude protein at 11.8 MJ ME/kg and not 12.8 or 13.8 MJ/kg. As dietary CP increased from 15 to 19%, breast meat yield increased by 10.8% on d 35 (P<0.01). Conversely, increasing metabolizable energy from 11.8 to 13.8 MJ/kg increased dressing percentage, breast skin, and subcutaneous fat, but decreased breast meat yield (% but not weight) on d 35 (P<0.01). In Experiment 2, the determined AMEn for diets formulated to contain 11.8, 12.8, or 13.8 MJ ME/kg were 11.66, 12.68, and 13.75 MJ/kg, respectively; determined standardized ileal digestible Lys was 0.95, 1.00, and 1.21% for diets formulated to contain 15, 17, or 19% crude protein, respectively. The best body weight gain and feed conversion ratio was obtained when ducks were fed a high dietary AMEn (13.75 MJ/kg) and high CP (19%, 1.21% SID Lys). These results provide a framework for subsequent modeling of amino acid and energy inputs and the corresponding outputs of growth performance and carcass components.

Effect of dietary methionine content on growth performance, carcass traits, and feather growth of Pekin duck from 15 to 35 days of age.

A study was conducted to establish the response of Pekin ducks to dietary Met from 15 to 35 d age. Experimental diets were formulated to contain 0.35, 0.45, 0.55, 0.65, and 0.75% Met (0.30, 0.39, 0.45, 0.56, and 0.68% on an analyzed basis, respectively) and 0.3% cysteine (0.25, 0.27, 0.26, 0.26, and 0.28% on an analyzed basis, respectively). Each diet was fed to 10 pens of 55 ducks/pen. Carcass yields and feather growth were determined at 28 and 35 d. Results showed that feeding 0.30% Met (0.55% Met+Cys) significantly impaired ADG, feed-to-gain (F:G) ratio, breast meat yield, and feather growth in comparison to the other dietary treatments (P < 0.05). BW, ADG, F: G, carcass and breast meat weight and yield, breast skin and subcutaneous fat weight and yield, the fourth primary wing feather length, and feather coverage showed significant quadratic broken-line or quadratic polynomial response to increasing dietary Met (P < 0.05). From 15 to 28 d age, the optimal Met requirement for the BW, breast meat yield, and the fourth primary wing feather length were 0.510, 0.445, and 0.404%, respectively, based on quadratic broken-line model, and correspondingly were 0.606, 0.576, and 0.559% by quadratic regression. For ducks from 15 to 35 d age, the optimal Met requirement for BW, breast meat yield, and feather coverage were 0.468, 0.408, and 0.484%, respectively, by quadratic broken-line model, and 0.605, 0.564, and 0.612%, by quadratic regression, respectively.

Dietary threonine requirement of Pekin ducks from 15 to 35 days of age based on performance, yield, serum natural antibodies, and intestinal mucin secretion.

A study was conducted to establish the dietary Thr requirement of Pekin ducks from 15 to 35 d of age. Experimental diets were formulated to contain 0.55, 0.60, 0.65, 0.75, and 0.85% Thr (0.57, 0.60, 0.64, 0.72, and 0.80% on an analyzed basis) and were studied in 2 experiments. In experiment 1, each diet was fed to 10 pens of 52 drakes per pen. Samples were collected at d 35 for determinations of carcass yields, serum immune parameters, and intestinal characteristics. Experiment 2 was a digestibility study, wherein 0.5% chromic oxide was mixed into the experimental diets and fed from 15 to 19 d. Ileal digesta were collected at d 19 to analyze mucin secretions and apparent ileal Thr digestibility. The results showed that feeding 0.72% versus 0.64% Thr improved 15 to 35 d BW gain by 55 g (P < 0.05), reduced feed-to-gain by 0.04 (P < 0.05), as well as increased carcass and breast meat yields by 22 and 24 g, respectively. Also, 0.72% Thr had the highest crude mucin secretion on a DM intake (DMI) basis (P < 0.05), although Thr had no effect on villus height, crypt depth, goblet cells, and MUC2 gene expression in the jejunum and ileum. In addition, serum natural IgY linearly increased (P < 0.0001) with dietary Thr increase. Using nonlinear regressions, Thr requirement was estimated to range from a low of 0.70% to maximize dry crude mucin secretion on a DMI basis to a high of 0.80% to maximize carcass weight and serum IgY production by the linear or quadratic regression. Equivalently, Thr requirement varied between a low of 0.62% to minimize mortality and a high of 0.73% to maximize dry crude mucin secretion expressed as DMI using the quadratic broken-line model. Correspondingly, the apparent ileal digestible Thr requirements were estimated to be 0.52 to 0.66% (0.70 to 0.80% dietary Thr) by quadratic and 0.47 to 0.56% (0.62 to 0.73% dietary Thr) by quadratic broken-line model.

A bovine herpesvirus type 1 mutant virus specifying a carboxyl-terminal truncation of glycoprotein E is defective in anterograde neuronal transport in rabbits and calves.

Bovine herpesvirus type 1 (BHV-1) is an important component of the bovine respiratory disease complex (BRDC) in cattle. The ability of BHV-1 to transport anterogradely from neuronal cell bodies in trigeminal ganglia (TG) to nerve ending in the noses and corneas of infected cattle following reactivation from latency plays a significant role in the pathogenesis of BRDC and maintenance of BHV-1 in the cattle population. We have constructed a BHV-1 bacterial artificial chromosome (BAC) clone by inserting an excisable BAC plasmid sequence in the long intergenic region between the glycoprotein B (gB) and UL26 genes. A BAC-excised, reconstituted BHV-1 containing only the 34-bp loxP sequence within the gB-UL26 intergenic region was highly infectious in calves, retained wild-type virulence properties, and reactivated from latency following treatment with dexamethasone. Using a two-step Red-mediated mutagenesis system in Escherichia coli, we constructed a gE cytoplasmic tail-truncated BHV-1 and a gE-rescued BHV-1. Following primary infection, the gE cytoplasmic tail-truncated virus was efficiently transported retrogradely from the nerve endings in the nose and eye to cell bodies in the TG of calves and rabbits. However, following dexamethasone-induced reactivation from latency, the gE mutant virus was not isolated from nasal and ocular sheddings. Reverse transcriptase PCR assays detected VP5 transcription in the TG of rabbits infected with gE-rescued and gE cytoplasmic tail-truncated viruses during primary infection and after dexamethasone treatment but not during latency. Therefore, the BHV-1gE cytoplasmic tail-truncated virus reactivated in the TG; however, it had defective anterograde transport from TG to nose and eye in calves and rabbits.

Immunohistochemical application of an antibody specific for human CD1a to the diagnosis of canine mast cell tumour.

Canine mast cell tumours (MCTs) may be graded microscopically for prognostic purposes. Grade I (well-differentiated) and grade II (intermediate differentiation) tumours have an abundance of metachromatic granules within the cytoplasm; however, grade III (poorly differentiated) MCTs may be difficult to diagnose as they frequently have fewer discernable granules. Herein we report that a cross-reactive anti-human CD1a monoclonal antibody (clone O10) may be used in immunohistochemistry to identify canine MCTs of all grades. The antibody was applied to tissue sections from 48 canine MCTs of different histological grades. Serial sections from each tumour were stained with toluidine blue and safranin O to compare diagnostic sensitivity. All MCTs were labelled positively by the CD1a antibody, but histochemical staining was often equivocal and identification of mast cells was extremely difficult in some cases. This antibody did not label neoplastic cells in cases of canine histiocytoma, plasmacytoma or amelanotic melanoma; therefore, the reagent may be a valuable marker for the diagnosis of canine MCTs, especially those tumours of histological grade III.

Analysis of bovine trigeminal ganglia following infection with bovine herpesvirus 1.

Following primary infection of the eye, oral cavity, and/or nasal cavity, bovine herpesvirus 1 (BHV-1) establishes latency in trigeminal ganglionic (TG) neurons. Virus reactivation and spread to other susceptible animals occur after natural or corticosteroid-induced stress. Infection of calves with BHV-1 leads to infiltration of lymphocytes in TG and expression of IFN-gamma (interferon-gamma), even in latently infected calves. During latency, virus antigen and nucleic acid positive non-neural cells were occasionally detected in TG suggesting there is a low level of spontaneous reactivation. Since we could not detect virus in ocular or nasal swabs, these rare cells do not support high levels of productive infection and virus release or they do not support virus production at all. Dexamethasone (DEX) was used to initiate reactivation in latently infected calves. Foci of mononuclear or satellite cells undergoing apoptosis were detected 6h after DEX treatment, as judged by the appearance of TUNEL+ cells (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling). BHV-1 antigen expression was initially detected in lymphocytes and other non-neural cells in latently infected calves following DEX treatment. At 24h after DEX treatment, viral antigen expression and nucleic acid were readily detected in neurons. Our data suggest that persistent lymphocyte infiltration and cytokine expression occur during latency because a low number of cells in TG express BHV-1 proteins. Induction of apoptosis and changes in cytokine expression following DEX treatment correlates with reactivation from latency. We hypothesize that inflammatory infiltration of lymphoid cells in TG plays a role in regulating latency.

Laboratory findings associated with abomasal ulcers/tympany in range calves.

The etiology of abomasal ulcers/tympany was investigated in 48 animals from 36 ranches in Wyoming and Nebraska. Results indicate that subclinical trace mineral deficiencies of copper and/or selenium exist in the range cattle in west central Nebraska and Wyoming. Etiological agents most frequently incriminated by bacteriologic cultures and/or histopathic examination were Clostridium perfringens and Campylobacter species. Histopathologic evaluation of abomasums revealed 31 of 38 cases contained abundant gram-positive bacteria associated with the damaged abomasal mucosa. Campylobacter-like organisms were demonstrated in 9 of 38 cases using the modified Dieterle stain. Clostridium perfringens was isolated in 14 of 38 cases, and Campylobacter jejuni was recovered from 5 of 38 cases.

Detection of canine parvovirus in naturally infected dogs with enteritis and myocarditis by in situ hybridization.

An improved method for the diagnosis of canine parvovirus using in situ hybridization in standard formalin-fixed, paraffin-embedded tissue sections was developed. A digoxigenin-labeled probe complementary to DNA sequences that code for the entire sequence of the capsid protein VP-1 and the middle part of the sequence of the capsid protein VP-2 was designed. Specific histologic localization of canine parvovirus-infected cells was demonstrated in small intestine, tonsil, lymph node, thymus, spleen, heart, liver, and kidney from dogs diagnosed at necropsy with canine parvovirus infection. The in situ hybridization accurately pinpointed the specific sites of viral infection. The detection of canine parvovirus in liver, kidney, and heart tissues together in the same pups could represent an enhanced virulence of this strain of canine parvovirus and suggests a broadened tissue tropism not seen before in Korean strains of canine parvovirus.

Porcine reproductive and respiratory syndrome: NEB-1 PRRSV infection did not potentiate bacterial pathogens.

A 2-phase study was conducted to evaluate the ability of the NEB-1 strain of porcine reproductive and respiratory syndrome virus (PRRSV) to potentiate common bacterial pathogens of swine. In phase I, 25 of 50 4-5-week-old specific-pathogen-free (SPF) pigs were exposed to NEB-1 PRRSV (day 0). Seven days after virus inoculation, 8 groups received 1 of 4 bacterial pathogens: Haemophilus parasuis, Streptococcus suis, Salmonella cholerasuis, and Pasteurella multocida. The ability of NEB-1 PRRSV to produce clinical disease, viremia, neutralizing antibody, gross and microscopic lesions and to potentiate bacterial pathogens was assessed. Response to NEB-1 PRRSV was similar among inoculated pigs; prolonged hyperthermia, lethargy, mild to moderate dyspnea, and cutaneous erythema were consistent clinical signs. No clinical differences were observed in groups after bacterial challenge. Virus was isolated from serum at weekly intervals through the end of the study, and all PRRSV-inoculated pigs had seroconverted by study termination. Two of 5 pigs died in non-PRRSV-inoculated groups challenged with H. parasuis and Streptococcus suis. Mortality in PRRSV-infected pigs was limited to 1 of 5 pigs from the Salmonella cholerasuis-challenged group. Gross lesions were seen in pigs dying after inoculation in H. parasuis- and Streptococcus suis-inoculated groups, in Salmonella cholerasuis- and P. multocida-challenged pigs, and in 1 non-PRRSV-inoculated control pig. Microscopic lesions consisted of mild to moderate proliferative interstitial pneumonia, nonsuppurative myocarditis, lymphoid hyperplasia, and nonsuppurative encephalitis in PRRSV-inoculated pigs. Findings in phase I indicated that NEB-1 PRRSV does not potentiate bacterial disease while inducing consistent clinical signs, viremia, seroconversion, and microscopic lesions.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of antibody response to porcine reproductive and respiratory syndrome virus ORF5 product following infection and evaluation of its diagnostic use in pigs.

The sensitivity and specificity of recombinant open reading frame 5 products used in the Western blotting assay for confirmation of porcine reproductive and respiratory syndrome virus (PRRSV) serologic status were evaluated. The recombinant antigen-based assays were specifically compared with a commercial enzyme-linked immunosorbent assay (ELISA) for PRRSV antibodies using 1) PRRSV antibody-negative reference sera (n = 30), 2) naturally infected pig sera (n = 40), 3) sequential sera obtained from 24 experimentally infected pigs, and 4) sera submitted to 3 state diagnostic laboratories (n = 200). The recombinant antigen assay yielded an average increased sensitivity of 10% over the commercial PRRSV ELISA. The negative controls (group 1 sera) showed no difference between the 2 assays. This comparison confirmed that the recombinant antigen-specific assay was more sensitive than the commercial ELISA and is well suited for routine confirmation of the presence of PRRSV antibodies.

Cryptosporidia in the cloacal coprodeum of red-lored parrots (Amazona autumnalis).

Cryptosporidiosis was diagnosed in the cloacal coprodeum of two red-lored parrots (Amazona autumnalis). The parasites were adhered to the microvillus border of epithelial cells and their presence was associated with loss and atrophy of microvilli. Merozoites, trophozoites, gametes, and oocytes were identified with light and transmission electron microscopy. The coprodeal epithelium was hyperplastic and the lamina propria was infiltrated with moderate numbers of heterophils and lymphocytes.

Liposarcoma in a Canada goose (Branta canadensis).

A multicentric liposarcoma was identified in various skeletal muscles of a mature Canada goose. The light and electron microscopic appearance of the tumor is discussed in relation to the appearance of liposarcoma in other species.

Seminoma and parathyroid adenoma in a snow leopard (Panthera unica).

A seminoma and parathyroid adenoma were diagnosed in an aged snow leopard. The ultrastructural appearance of the seminoma was similar to that described in the dog and in man. The lack of significant amounts of rough endoplasmic reticulum, Golgi complexes and free ribosomes in the parathyroid adenoma suggested that it was non-functional. Parathyroid adenoma has not been previously described in a large wild feline.

Identification of Mycoplasma hyopneumoniae in formalin-fixed porcine lung, using an indirect immunoperoxidase method.

An indirect immunoperoxidase staining technique was evaluated for detection of Mycoplasma hyopneumoniae in formalin-fixed, paraffin-embedded porcine lung. Lungs from swine with induced (n = 4) or naturally occurring M hyopneumoniae infection (n = 31) were examined grossly, by light and immunofluorescent microscopy, and by an indirect immunoperoxidase test, using antibody raised in swine against M hyopneumoniae as the primary antibody. Organisms stained by the indirect immunoperoxidase method were identified in tissue sections as pleomorphic brown-staining structures corresponding to those observed with immunofluorescence. Mycoplasma hyosynoviae, M hyorhinis, and Acholeplasma laidlawii did not stain with the indirect immunoperoxidase method, using antibody raised against M hyopneumoniae.

Long-term effect of Bordetella bronchiseptica infection in neonatal pigs.

A total of 62 gnotobiotic pigs were inoculated with Bordetella bronchiseptica at 7 days of age. There were 25 noninoculated controls. Thirteen of the inoculated pigs were killed and 6 pigs which died were examined between 3 and 5 weeks after inoculations were done (group A). Of these 19 pigs, 16 had severe pneumonia and B bronchiseptica was isolated from the lungs of 17. The turbinates in most of the pigs appeared to have some atrophy. Four other pigs died of pneumonia after they were transferred to the floor in isolation rooms, and 9 inoculated pigs were examined at necropsy at about 112 days of age (group B). Bordetella bronchiseptica was isolated from 8 of the latter 9 pigs, and 7 of the pigs had pneumonic lesions. The turbinates all appeared normal. Thirty inoculated pigs (group C) were killed at 180 days of age. Of these, 22 had pneumonic lesions. Bordetella bronchiseptica was reisolated from the turbinates of 2 of 30 pigs, although the turbinates in all appeared normal. The noninoculated controls examined at necropsy at 3 to 5 weeks, 112 days, and 180 days of age did not have any pulmonic lesions, and their turbinates appeared normal. The inoculated pigs killed at 112 days of age gained an average of 0.25 kg/day, and the control pigs gained 0.31 kg. At 180 days of age, the inoculated pigs had gained 0.41 kg/day and the controls, 0.49 kg. Bordetella bronchiseptica infection in susceptible pigs can cause a severe pneumonia which can persist and reduce the growth rate.

Pathophysiologic studies of calves given 3-methylindole intraruminally.

Intraruminal administration of 0.25 g of 3-methylindole (3MI; skatole/kg of body weight) to seven young calves generally caused mild respiratory signs and lesions, accompanied by only slight changes in cardiopulmonary function. Moderate depression, trembling, and irregular respiratory rate were observed between postadministration hours (PAH) 6 and 12. By PAH 24 at this dosage, abnormal clinical signs were not present. Statistically significant (P less than or equal to 0.05) changes observed in blood gas data from the seven calves were a decrease in aortic oxygen tension at PAH 12, increases in free-flowing venous oxygen tension in the intervals between PAH 6 and 12 and between PAH 6 and 24, and an increase in occluded venous oxygen tension at PAH 24. All calves had increases (although generally not statistically significant) in heart rate, cardiac output, cardiac index, stroke volume, and stroke index after 3MI administration. Mean aortic and pulmonary arterial pressure changes were generally small and variable. At necropsy, the lungs of the calves did not collapse when the thorax was opened. Patchy areas of consolidation (0.5 cm in diameter) were scattered throughout the parenchyma. Pulmonary edema or emphysema was not observed grossly. Microscopically, the alveolar septae were irregularly thickened because of edema, infiltration by polymorphonuclear and mononuclear cells, and vascular congestion. Interstitial lesions were patchy in distribution and severity and corresponded to the areas of consolidation observed grossly. Alveolar epithelial hypertrophy and hyperplasia were present, and an occasional focus of alevoli contained fluid of edema. Degeneration of individual hepatocytes was observed in scattered areas of the liver, especially in the periportal areas. It was concluded that differences in 3MI dosage response may exist between young calves and adult cattle in which calves are more resistant to the pulmonary cytotoxicity of 3MI.

Comparative pathogenicity of selected bovine viral diarrhea virus isolates in gnotobiotic lambs.

The infectivity and pathogenicity of selected bovine viral diarrhea virus (BVDV) isolates were determined in gnotobiotic, colostrum-deprived neonatal lambs. Five-day-old cesarean-derived gnotobiotic lambs were exposed to 1 of 10 BVDV isolates via aerosol suspension. These isolates were from tissues or secretions of calves or lambs affected with respiratory tract disease, weak neonatal calves, aborted bovine fetuses, or reference Singer or Draper BVDV. The pathogenicity of each isolate, relative to the others, was evaluated in lambs by measurement of the neutralizing antibody response, virus isolation from nasal secretions or tissues, and postmortem lesions. The BVDV isolates varied in their infectivity and pathogenicity. Singer, the cytopathic reference strain, was the most lymphotrophic isolate and stimulated the greatest neutralizing antibody response. Encephalitis was the most consistent lesion observed and was used as the final determinant of relative pathogenicity of the viruses. The most neuropathogenic isolates were the 2 viruses originating from lambs affected with respiratory tract disease, the 2 weak neonatal calf isolates, and 1 isolate from an aborted bovine fetus. The least pathogenic isolates were the 2 reference isolates, Draper and Singer; the 2 mucosal disease isolates; and 1 isolate originating from an aborted bovine fetus.

Isolation of Streptococcus suis from swine in Nebraska.

Streptococcus suis was isolated from swine with localized and diffuse infections. The isolates (n = 121) were unable to grow in broth containing 6.5% NaCl. Their abilities to hydrolyze esculin were variable, as were their fermentative reactions in lactose, trehalose, raffinose, and inulin. Sorbitol was not attacked by any strain. Serologic reactions were obtained between the isolates and coagglutination reagents prepared from commercially available antisera to streptococcal groups D, R, or S.

Trichosporonosis in two cats.

Infection with Trichosporon spp was diagnosed in 2 cats. In one cat, infection consisted of a granulomatous dermatitis and was concurrent with disseminated lymphoblastic lymphosarcoma. In another cat, urinary cystitis caused by T beigelii was diagnosed.