Metabolic disorder of amino acids, fatty acids and purines reflects the decreases in oocyte quality and potential in sows.

Oocyte quality is closely related to female fertility. Nevertheless, core nutritional metabolites influencing oocyte quality are unclear. Herein, comprehensive metabolomics analysis of follicular fluid, serum, and urine from low reproductive performance (LRP) and normal reproductive performance (NRP) sows was conducted. Twenty-seven, fourteen and sixteen metabolites (involved in metabolism of amino acids, fatty acids, purine and pyrimidine) were altered in follicular fluid, serum and urine, respectively, in LRP compared with NRP sows, and could decrease oocyte quality and developmental potential, ultimately leading to low fertility. Deoxyinosine, guanidine acetate, thymidine, 5,6-epoxy-eicosatrienoic acid, carnosine, docosahexaenoic acid and carbamoyl phosphate in follicular fluid, cysteine, carnitine, serotonin, hypoxanthine, valine and arginine in serum, as well as carnitine, phenyl glycine, N-acetyl glutamine, propionyl carnitine and choline in urine could be selected as diagnostic markers to indicate oocyte quality. Consistent with metabolomics data, we confirmed changes in concentrations of fatty acids and amino acids in follicular fluid. Targeting purine metabolism, elevating levels of deoxyinosine in in-vitro maturation medium of porcine oocyte significantly promoted the blastocyst rate. Collectively, this study provided new information of potential targets for predicting oocyte quality and developmental potential, and may help with strategies for early diagnosis or therapeutic/dietary intervention in improving reproductive outcomes.

Genetic diversity, virulence genotype and antimicrobial resistance of uropathogenic Escherichia coli (UPEC) isolated from sows.

BACKGROUND: Urinary tract infections (UTI) cause severe losses to the swine industry worldwide and uropathogenic Escherichia coli (UPEC) are the main agent isolated from UTI in sows. OBJECTIVE: The aim of this study was to investigate the virulence genes, assess the phylogenetic background, clonal diversity, and the pattern of resistance to antimicrobials in 186 isolates of UPEC isolated from sows in Brazil. MATERIALS AND METHODS: Urine samples from 300 sows of three herds with clinical signs from São Paulo State (Brazil) were screened for UTI; samples with suggestive results were submitted to bacterial isolation. E. coli strains isolated were characterized using disk diffusion technique, polymerase chain reaction and Single-enzyme amplification fragment length polymorphism (SE-AFLP). RESULTS: Virulence genes focH and papC were present in 78.5% and 58% of strains, respectively, followed by cnf1 (23.2%), afa (13.4%), sfa (11.3%), iucD (6.9%), and hlyA (1.6%). No clonal relatedness was found by SE-AFLP. A total of 98% of isolates (182/186) were multidrug resistant, and the highest levels of resistance were to sulfonamides, tetracycline, florfenicol, and ampicillin. Isolates were classified in phylogenetic group B1 (34.4%), followed by D (33.9%), E (30.1%) and A (1.6%). CONCLUSIONS: The data obtained suggest that pigs from clinically affected herds may serve as a reservoir of uropathogenic and multidrug-resistant E. coli strains.

Acute encephalopathy in a 2-year-old pot-bellied pig following accidental intoxication with clonazepam.

OBJECTIVE: To describe a case of successful management of clonazepam toxicity causing encephalopathy in a pot-bellied pig. CASE SUMMARY: A 2-year-old female pot-bellied pig weighing 13.5 kg was presented for evaluation of clinical signs of acute encephalopathy. Based on the animal's history and clinical signs, a tentative diagnosis of benzodiazepine (BZP) intoxication was made. The results of a urinary drug screening test designed to detect illicit substances in human urine indicated benzodiazepine exposure. Gas chromatography and mass spectrometry analysis later confirmed clonazepam (urinary concentration 496 ng/mL) as the intoxicating substance. The pig responded favorably to treatment which included administration of flumazenil, decontamination with enteral activated charcoal, and intravenous isotonic crystalloid administration. The pig had a rapid improvement in mentation 10 minutes following IV flumazenil administration and was considered mentally appropriate following 24 hours of hospitalization. The pig was discharged from the hospital after 48 hours of care, and was reported to be doing well 6 months later. NEW INFORMATION PROVIDED: Intoxication with prescription benzodiazepines can occur in companion animals and result in clinical signs of acute encephalopathy. Urinary drug screening tests designed for human use may provide rapid results to indicate drug intoxication and guide therapeutic intervention in veterinary species. Administration of flumazenil resulted in a rapid improvement in mentation following clonazepam intoxication in a pot-bellied pig.

Urinary Hepcidin Levels in Iron-Deficient and Iron-Supplemented Piglets Correlate with Hepcidin Hepatic mRNA and Serum Levels and with Body Iron Status.

Among livestock, domestic pig (Sus scrofa) is a species, in which iron metabolism has been most intensively examined during last decade. The obvious reason for studying the regulation of iron homeostasis especially in young pigs is neonatal iron deficiency anemia commonly occurring in these animals. Moreover, supplementation of essentially all commercially reared piglets with iron entails a need for monitoring the efficacy of this routine practice followed in the swine industry for several decades. Since the discovery of hepcidin many studies confirmed its role as key regulator of iron metabolism and pointed out the assessment of its concentrations in biological fluids as diagnostic tool for iron-related disorder. Here we demonstrate that urine hepcidin-25 levels measured by a combination of weak cation exchange chromatography and time-of-flight mass spectrometry (WCX-TOF MS) are highly correlated with mRNA hepcidin expression in the liver and plasma hepcidin-25 concentrations in anemic and iron-supplemented 28-day old piglets. We also found a high correlation between urine hepcidin level and hepatic non-heme iron content. Our results show that similarly to previously described transgenic mouse models of iron disorders, young pigs constitute a convenient animal model to explore accuracy and relationship between indicators for assessing systemic iron status.

Urinary carbonic anhydrase VI as a biomarker for kidney disease in pigs.

This study investigated whether carbonic anhydrase (CA)-VI has utility as a biomarker in swine kidney disease. Serum chemistry, histopathology, immunohistochemical staining and enzyme-linked immunosorbent assay (ELISA) analyses were performed. In the kidney of normal healthy pigs, CA-VI was localized in the epithelial cells of the renal distal straight tubules. CA-VI levels were 16 ± 35 ng/g wet tissue and 50 ± 66 ng/mL in normal pig kidney and urine, respectively, and 136 ± 173 ng/mL in the urine of pigs with kidney disease. CA-VI urinary concentration was not correlated with urinary urea nitrogen (UUN), urinary creatinine (Cre), or urinary albumin levels in pigs with kidney disease. However, UUN and Cre levels were positively correlated in the urine of pigs with kidney disease. These data suggest that urinary CA-VI may represent a biomarker for kidney disease in pigs, particularly for disorders affecting distal straight tubules.

Identification of factors associated with increased excretion of foot-and-mouth disease virus.

We investigated which variables possibly influence the amount of foot-and-mouth disease virus (FMDV) shed in secretions and excretions by FMDV infected animals, as it is likely that the amount of FMDV shed is related to transmission risk. First, in a separate analysis of laboratory data, we showed that the total amount of FMDV in secretions and excretions from infected animals is highly correlated with maximum titres of FMDV. Next, we collected data from 32 published scientific articles in which FMDV infection experiments were described. The maximum titres of FMDV reported in different secretions and excretions (the response variable) and the experimental conditions in which they occurred (the explanatory variables), were recorded in a database and analyzed using multivariate regression models with and without random effects. In both types of models, maximum titres of FMDV were significantly (p<0.05) associated with types of secretions and excretions, animal species, stage of the disease and days post infection. These results can be used to prioritize biosecurity measures in contingency plans.

Urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock.

The aim of the present study was to consider the wide usage of urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock. A total of 512 adult animals (300 cattle, 138 horses, 59 goats and 15 pigs), from herds/flocks with reproductive problems in Rio de Janeiro, Brazil was studied by serology and urinary PCR. From the 512 serum samples tested, 223 (43.5 %) were seroreactive (cattle: 45.6 %, horses: 41.3 %, goats: 34%and pigs: 60 %). PCR detected leptospiral DNA in 32.4 % (cattle: 21.6 %, horses: 36.2 %, goats: 77.4 % and pigs: 33.3 %. To our knowledge there is no another study including such a large number of samples (512) from different species, providing a comprehensive analysis of the usage of PCR for detecting leptospiral carriers in livestock. Serological and molecular results were discrepant, regardless the titre, what was an expected outcome. Nevertheless, it is impossible to establish agreement between these tests, since the two methodologies are conducted on different samples (MAT - serum; PCR - urine). Additionally, the MAT is an indirect method and PCR is a direct one. In conclusion, we have demonstrated that urinary PCR should be considered and encouraged as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock.

Actinobaculum suis detection using polymerase chain reaction.

Actinobaculum suis is an important agent related to urinary infection in swine females. Due to its fastidious growth characteristics, the isolation of this anaerobic bacterium is difficult, thus impairing the estimation of its prevalence. The purpose of this study was to develop and test a polymerase chain reaction (PCR) for the detection and identification of A. suis and then compare these results with traditional isolation methods. Bacterial isolation and PCR were performed on one hundred and ninety-two urine samples from sows and forty-five preputial swabs from boars. The results indicate that this PCR was specific for A. suis, presenting a detection limit between 1.0 × 10(1) CFU/mL and 1.0 × 10(2) CFU/mL. A. suis frequencies, as measured by PCR, were 8.9% (17/192) in sow urine samples and 82.2% (37/45) in preputial swabs. Assessed using conventional culturing techniques, none of the urine samples were positive for A. suis; however, A. suis was detected in 31.1% (14/45) of the swabs. This PCR technique was shown to be an efficient method for the detection of A. suis in urine and preputial swabs.

Urinary excretion of collagen degradation markers by sows during postpartum uterine involution.

Incomplete uterine involution is the putative cause of the increased embryo mortality and reproductive failure often exhibited by sows that lactate for less than 21 days. Since such short lactation lengths are common in American swine production, an effective technique to monitor the postpartum involution process and test this hypothesis might be valuable. Rapid and extensive catabolism of uterine collagen is essential for normal postpartum involution. The objective of this study was to characterize postpartum excretion of two biochemical markers of collagen degradation. In experiment I, urine samples were collected from five sows every other day from the day before parturition (day -1), through a 21-day lactation, to day 8 postweaning. The collagen crosslinks hydroxylysyl pyridinoline (HP), which is present in many tissues, and lysyl pyridinoline (LP), which is primarily concentrated in bone, were assayed by both ELISA and HPLC. Urinary levels of both free (ELISA) and total (HPLC) HP and LP increased (P < 0.001) approximately two-fold during lactation. The mean molar ratio of total HP:LP increased (P < 0.001) from 6.6 +/- 1.6 at day 1 to a maximum of 10.2 +/- 1.5 at day 7 postpartum and averaged 9.1 +/- 0.3 for the entire sampling period. These data are consistent with a postpartum increase of soft tissue collagen catabolism since bone has a low HP:LP ratio of 4 and soft tissues like the uterus have a high HP:LP ratio of >/=20 because they contain only trace amounts of LP. Since HPLC (total) and ELISA (free) crosslinks estimates were highly correlated (r = 0.85-0.91, P < 0.001) in experiment I, only the less technical ELISA technique was used in experiment II. Urine samples were collected from 21 sows every third day from day 1 to 19 of lactation. Sows from this second group exhibited one of four distinct crosslinks excretion patterns: peak on day 1 (n = 3), peak on day 7 (n = 4), peak on day 10, 13 or 16 (n = 7), or no peak (n = 7). This variation of postpartum crosslinks excretion among sows was not related to parity, body weight, lactation body weight change, litter size, or litter birth weight. Overall, data from experiments I and II indicate that urinary HP does increase postpartum in a pattern temporally consistent with uterine involution. However, significant variation among sows in the magnitude and timing of peak HP excretion was evident.

Metabolic profile of pigs fed feed containing zearalenone destructor.

Zearalenone (ZEA) is a member of macrocyclic lactons family. It is a toxin--phytosteride produced by fungi of Fusarium ssp. genus. Zearalenone contaminates food and animal feeding stuffs and its destruction is difficult. It requires application of particular compounds that would bind zearalenone in the feed or feeding stuff or in the gastrointestinal tract and decrease its bio-accessibility. It should also fulfil all the safety requirements regarding the plant supplements and animals that are fed with this feed. The aim of the study was to estimate if the feed supplemented with different doses of zearalenone and zearalenone destructor causes changes of the metabolic profile in gilts. The results obtained show that applied destructor did not cause negative haematological and biochemical changes in the blood of the gilts examined. It can be suggested that it is a safe feed supplement pigs in prevention of zearalenone micotoxicosis.

Osteopenic disease in growing pigs: diagnostic methods using serum and urine calcium and phosphorus values, parathoromone assay, and bone analysis.

This research was performed to evaluate the utility of several serum and urine parameters as well as bone ash and plasma parathormone assay to diagnose and monitor diet-related osteopenia in growing pigs. Five diets were tested as follows: calcium-deficient, phosphorus-replete; moderate-deficiency of calcium and phosphorus; marked deficiency of calcium and phosphorus; calcium replete, phosphorus deficient; and vitamin D deficient. Parameters monitored included serum calcium and phosphorus as well as ratios of urine calcium to creatinine, phosphorus to creatinine, calcium to phosphorus, and percent fractional excretions of calcium and phosphorus. Plasma parathormone (PTH) levels were monitored in 2 of 3 experiments. Osteopenic bone differences at necropsy were evaluated by bone density, percent ash, ash per milliliter bone, calcium per milliliter bone, and phosphorus per milliliter bone. Marked change in urine mineral parameters, especially the calcium-to-phosphorus ratio, typically occurred within 1 to 2 days of treatment and preceded significant change in serum mineral or plasma PTH by 2 to 3 weeks. When monitored, plasma PTH levels were elevated following treatment, which confirms the hyperparathyroid state induced by the test diets. Significant differences in bone mineralization between control and treatment diets at necropsy were generally observed. The results of this study indicate that the analysis of urine minerals offers an early, noninvasive technique to investigate diet-associated osteopenic disease in growing pigs, which can be supported further by bone mineral analysis at postmortem using techniques herein described. Several urine mineral reference intervals for application to field investigations are included. Research into application of similar techniques to evaluate calcium and phosphorus homeostasis in pigs of all ages, including gestating and lactating gilts and sows, appears warranted.

Doramectin efficacy against the kidney worm, Stephanurus dentatus in sows.

Sixteen sows passing Stephanurus dentatus eggs in their urine were bought on the local market and placed in individual pens with solid concrete floors in an open-sided bam. Water was supplied by nipple waterers and sows were fed individually 1.8 kg feed daily. First, urine samples were taken at dawn on days -8 and -7 and weights were taken on day 0. Sows were assigned to one of two groups on the basis of average urine egg counts and weights. Group T1 sows were each injected IM in the neck with sterile saline at the rate of 1.5 ml per 50 kg and group T2 sows were each injected IM with doramectin at the rate of 300 micrograms kg-1 on day 0. Urine samples were again taken on days 56 and 57 and the sows were necropsied on day 57. Urine of all doramectin treated sows were test negative for kidney work eggs on days 56 and 57 as was one control sow, whereas the average count for controls was 3762 eggs ml-1. No worms were found in doramectin treated sows and a total of 499 were found in the controls for an average of 62 per sow. The majority of worms were in the perirenal area and kidneys, a few were scattered in liver, lungs, abdominal muscles and peritoneal cavity. The efficacy of doramectin against Stephanurus dentatus in sows was 100% (P < 0.0001).

[The peripartum disease complex of the sow in industrial swine breeds. 1. Peripartum course of bacteriuria of sows with vaginal-vulvar discharge in a modern swine breeding facility]. / Bericht über den peripartalen Krankheitskomplex der Muttersau in der industriellen Schweinezucht. 1. Mitteilung: Peripartaler Verlauf der Bakteriurie der Sauen mit vaginal-vulvalem Ausfluss in einem modernen Schweinezuchtbetrieb.

In an industrial pig production unit ten sows were selected at random. All the sows had puerperal disease in their anamnesis and all of them revealed at the time of selection--during their late pregnancy--Urinary Tract Infection (UT)I and vaginal-vulvar discharge (VD). Mid-stream early morning urine samples were collected during four days ante partum and during four days post partum. The samples were semiquantitatively examined for E. coli and gram positive cocci. Three sows showed ante partum non significant bacteriuria which turned into significant bacteriuria post partum. The majority of the reminding sows revealed a significant bacteriuria during the whole period of examination.

Rapid and specific detection of pathogenic Leptospira species by amplification of ribosomal sequences.

We have developed an assay for the detection of pathogenic Leptospira that is based on the polymerase chain reaction. With the combination of agarose gel electrophoresis and blotting, pathogenic Leptospira can be discriminated specifically from nonpathogenic Leptospira and other bacterial species. This method, based on the amplification of 16S ribosomal RNA sequences, is able to detect 10 leptospiral cells/mL in cattle urine samples and 100 leptospiral cells/mL in pig urine samples. Using this assay leptospires were detected in urine samples from cattle that were experimentally infected with Leptospira interrogans serovar hardjo type hardjobovis.

[Pteridine pattern in the urine of swine of different breeds and halothane genotypes in relation to nutrition and stress]. / Untersuchungen zum Pteridinmuster im Harn von Schweinen verschiedener Rassen und Halothan-Genotypen in Abhängigkeit von Ernährung und Belastung.

Urinary pteridine patterns and creatinine concentrations were determined in pigs of the breeds Piétrain, Deutsche Landrasse and their crossbreds. Also the influence of the halothane genotypes, of different feeding and of treadmill exercise was examined. We conclude that creatinine concentration in pig urine is not a reliable reference parameter for the pteridine excretion. Creatinine values were significantly influenced by breed and feeding. The pteridine pattern of pigs' urine depends on the breed and correlates with other metabolic parameters. Stress is reflected by altered pteridine concentrations.

[The usefulness of urine analysis for the diagnosis of macroscopic lesions of the urogenital system of sows during slaughter]. / De bruikbaarheid van urine-onderzoek voor het opsporen van macroscopische afwijkingen aan het urogenitaalapparaat van zeugen bij het slachten.

As part of a research project concerned with the monitoring of health problems in sows, the practicability of post-mortem urinalysis of sows during slaughter was studied to determine the extent to which this analysis was useful. Samples of urine were taken from 232 sows after slaughter to test them for pH, nitrite and blood concentrations by test strips (Combur-8, Boehringer Mannheim). Positive results of testing were found not to be associated with pathological disorders of the genito-urinary system to any marked extent. Samples of urine taken pre- as well as post-mortem from 111 other sows were subjected to same tests. During the process of slaughter, the pH was found to show a decrease, whereas the concentrations of protein and blood increased. The results of post-mortem tests for nitrite were definitely not correlated with those of pre-mortem tests. The results of testing urine sampled prior to slaughter at the farm were related to post-mortem pathological findings to a greater extent than were the results of testing after slaughter.

The influence of nutritional factors on biopterin excretion in laboratory animals.

Nutritional influences on urinary total biopterin levels in rats and pigs were investigated. During the first nights in metabolic cages with food deprivation a significant increase in biopterin values was found in rats. This could be diminished either by feeding, adaptation to food deprivation or by oral glucose application. With food deprivation under normal housing conditions, this increase could not be found. Rats that were fed a cellulose preparation without metabolizable energy had no increase in biopterin excretion. The circadian rhythm of biopterin excretion was influenced by food deprivation as well as by cellulose. Alterations in water intake and urinary output had no effect on biopterin levels related to creatinine. Remarkable changes in biopterin excretion are more likely due to hormonal functions and regulations related to stress than to nutritional factors. More investigations into these problems are being performed.

[Corynebacterium suis infection in swine. 1. Clinical diagnosis with special consideration of urine studies and cystoscopy]. / Corynebacterium-suis-Infektion beim Schwein. 1. Mitteilung: Klinische Diagnose unter besonderer Berücksichtigung von Harnuntersuchung und Zystoskopie.

Experimental and spontaneous infections with Corynebacterium suis in sows were investigated. In early stages animals show no clinical disorders or only for a short time. However, there are already marked changes in urinary samples (hematuria, proteinuria, leukocyturia, gross alterations). Using an endoscope mucosal irritations can be seen mainly on the floor of the bladders. In chronic cases alterations in urine are more pronounced. If a pyelonephritis is present in addition to the cystitis, general signs of illness are evident including anorexia, emaciation, anemia, subnormal body temperature and abortions. Bladders demonstrate an erosive and ulcerative, hemorrhagic cystitis on the whole mucosal surface. Uremia appears only in late stages of the disease.