In vitro effect of Aloe vera, Coriandrum sativum and Ricinus communis fractions on Leishmania infantum and on murine monocytic cells

In South America, visceral leishmaniasis is a zoonosis caused by the protozoan species Leishmania infantum (syn. L. chagasi) and is primarily transmitted through the bite of the female Lutzomyia longipalpis. Its main reservoir in urban areas is the dog. The application of control measures recommended by health agencies have not achieved significant results in reducing the incidence of human cases, and the lack of effective drugs to treat dogs resulted in the prohibition of this course of action in Brazil. Therefore, it is necessary to search new alternatives for the treatment of canine and human visceral leishmaniasis. The objectives of this study were to evaluate the in vitro effect of fractions from Aloe vera (aloe), Coriandrum sativum (coriander), and Ricinus communis (castor) on promastigotes and amastigotes of L. infantum and to analyze the toxicity against the murine monocytic cells RAW 264.7. To determine the viability of these substances on 50% parasites (IC50), we used a tetrazolium dye (MTT) colorimetric assay (bromide 3-4.5-dimethylthiazol-2-yl-2,5-dephenyltetrazolium), and on amastigotes we performed an in situ ELISA. All fractions were effective against L. infantum promastigotes and did not differ from the positive control pentamidine (p>0.05). However, the R. communis ethyl acetate and chloroform fractions, as well as the C. sativum methanol fraction, were the most effective against amastigotes and did not differ from the positive control amphotericin B (p>0.05). The R. communis ethyl acetate fraction was the least toxic, presenting 83.5% viability of RAW 264.7 cells, which was similar to the results obtained with amphotericin B (p>0.05). Based on these results, we intend to undertake in vivo studies with R. communis ethyl acetate fractions due the high effectiveness against amastigotes and promastigotes of L. infantum and the low cytotoxicity towards murine monocytic cells.     

Campylobacter jejuni is associated with, but not sufficient to cause vibrionic hepatitis in chickens

Vibrionic hepatitis is a disease of poultry which is characterised by the presence of focal lesions in the liver, usually 1-2mm in size and greyish-white in colour. The cause of the disease remains unclear, as do the reasons for its recent re-emergence. We examined the livers of commercial broiler chickens taken during processing and found Campylobacter spp. in both normal livers and those displaying signs indicative of focal hepatitis. Livers with signs of hepatitis had significantly more Campylobacter spp. present than those without and other bacterial genera were infrequently present. We were unable to replicate the disease in a healthy host following experimental infection with a Campylobacter jejuni strain isolated from a liver showing signs of focal hepatitis. However, a significant T cell response to C. jejuni was seen in the liver of Campylobacter infected birds. We conclude that the presence of Campylobacter spp. in the liver alone is not sufficient to cause vibrionic hepatitis, but that a predisposing factor, possibly within the host is required. We also provide evidence that chickens mount an adaptive T cell response to systemic C. jejuni.     

Phenotypic and genotypic characterization of β-lactam resistance in Klebsiella pneumoniae isolated from swine

Little is known about the antimicrobial resistance mechanisms in Klebsiella pneumoniae from swine in China. Thus, this paper aims to demonstrate the β-lactam resistance phenotypes and genotypes of K. pneumoniae isolates from swine in southwestern China, detect possible new β-lactamase variants, and determine whether or not the variants differ in their antibiotic resistance. Isolates from 58 unrelated diseased swine were collected from 61 pig farms in southwestern China from 2007 to 2009. Among the 58 isolates, 75.8-100% were resistant to β-lactam, 62.0-68.97% to fluoroquinolone, 44.8-46.55% to aminoglycoside, and 8.62-17.24% to β-lactam inhibitors. PCR amplification and DNA sequencing showed that bla(TEM-1) was detected in 100% (n=58) of the isolates, bla(SHV) in 82.76% (n=48), bla(CTX-M) in 39.66% (n=23), and bla(OKP) in 17.24% (n=10). The bla(SHV) types included bla(SHV-1), bla(SHV-11), bla(SHV-12), and bla(SHV-27). None of the isolates harbored bla(KPC), bla(LEN), or bla(GES) gene. Four novel variants (bla(OKP-A-13), bla(OKP-A-14), bla(OKP-A-15), and bla(OKP-A-16)) were identified among the 10 OKP β-lactamase-producing K. pneumoniae isolates resistant to ampicillin, amoxicillin, oxacillin, cefalexin, and cefadroxil. Plasmid analysis and PCR amplification indicated that bla(TEM-1) genes were detected in the total plasmid. Molecular typing by pulsed-field gel electrophoresis revealed the presence of 10 distinct pulsotypes of OKP producer isolates. Plasmid DNA digested with XbaI yielded two to six bands of ca. 0.15-30 kb. Transformants of the 10 OKP producer isolates showed no differences in their antibiotic susceptibility, except for the pulsotype B transformant, which carried bla(CTX-M). In China, β-lactam resistance appeared to be common among K. pneumoniae isolates from swine, suggesting that K. pneumoniae may be a reservoir for the dissemination of β-lactam resistance among Chinese pig farms.     

Feruloylated and nonferuloylated arabino-oligosaccharides from sugar beet pectin selectively stimulate the growth of Bifidobacterium spp. in human fecal in vitro fermentations

The side chains of the rhamnogalacturonan I fraction in sugar beet pectin are particularly rich in arabinan moieties, which may be substituted with feruloyl groups. In this work the arabinan-rich fraction resulting from sugar beet pulp based pectin production was separated by Amberlite XAD hydrophobic interaction and membrane separation into four fractions based on feruloyl substitution and arabino-oligosaccharide chain length: short-chain (DP 2-10) and long-chain (DP 7-14) feruloylated and nonferuloylated arabino-oligosaccharides, respectively. HPAEC, SEC, and MALDI-TOF/TOF analyses of the fractions confirmed the presence of singly and doubly substituted feruloylated arabino-oligosaccharides in the feruloyl-substituted fractions. In vitro microbial fermentation by human fecal samples (n = 6 healthy human volunteers) showed a selective stimulation of bifidobacteria by both the feruloylated and the nonferuloylated long-chain arabino-oligosaccharides to the same extent as the prebiotic fructo-oligosaccharides control. None of the fractions stimulated the growth of the potential pathogen Clostridium difficile in monocultures. This work provides a first report on the separation of potentially bioactive feruloylated arabino-oligosaccharides from sugar beet pulp and an initial indication of the potentially larger bifidogenic effect of relatively long-chain arabino-oligosaccharides as opposed to short-chain arabino-oligosaccharides.     

Mulberry water extracts possess an anti-obesity effect and ability to inhibit hepatic lipogenesis and promote lipolysis

Obesity plays a critical role in dyslipidemia and related disorders. Mulberry water extracts (MWEs) contain polyphenols, including gallic acid, chlorogenic acid, rutin, and anthocyanins. In this study, using 6-week-old male hamsters, we investigated the anti-obese effect of MWEs. After 12 weeks of treatment, MWEs lowered high-fat diet (HFD)-induced body weight and visceral fat, accompanied with hypolipidemic effects by reducing serum triacylglycerol, cholesterol, free fatty acid, and the low-density lipoprotein (LDL)/high-density lipoprotein (HDL) ratio (n=8 for each group). MWEs decreased hepatic lipids, thus protected livers from impairment. The hepatic peroxisome proliferator-activated receptor α and carnitine palmitoyltransferase-1 were elevated, while fatty acid synthase and 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase were reduced by MWEs, indicating that MWEs regulated lipogenesis and lipolysis, which exerted the anti-obese and hypolipidemic effects. Noticeably, MWEs showed both efficacy and safety in vivo. In concluson, MWEs can be used to reduce body weight, serum, and liver lipids.     

Multiple immunofluorescence labelling enables simultaneous identification of all mature fibre types in a single equine skeletal muscle cryosection

Skeletal muscle is composed of a heterogeneous mixture of several fibre types, each with specific physiological properties. In equine muscle, identification of these individual fibres (fibre typing) is important for both exercise physiology and pathological studies. Traditionally, fibre typing has been achieved by adenosine triphosphatase (ATPase) histochemistry or by immunoperoxidase labelling with antibodies directed at myosin heavy chain isoforms. ATPase histochemistry can be temperamental and lacks specificity, and both techniques require staining of serial cryosections to reveal the entire fibre type compliment of a single sample, which is time consuming and prone to inaccuracy. Here we describe an immunofluorescence labelling technique that enables rapid, accurate and specific identification of the 3 mature equine muscle fibre types in a single cryosection.     

Detection of protective antibody titers against feline panleukopenia virus, feline herpesvirus-1, and feline calicivirus in shelter cats using a point-of-care ELISA

Serum antibody titers are a useful measurement of protection against infection (feline panleukopenia virus [FPV]) or clinical disease (feline herpesvirus-1 [FHV] and feline calicivirus [FCV]), and their determination has been recommended as part of disease outbreak management in animal shelters. The objective of this study was to determine the sensitivity, specificity, and inter-observer and inter-assay agreement of two semi-quantitative point-of-care assays for the detection of protective antibody titers (PAT) against FPV, FHV and FCV in shelter cats. Low sensitivity for FPV antibodies (28%) rendered a canine point-of-care assay inappropriate for use in cats. The feline point-of-care assay also had low sensitivity (49%) and low negative predictive value (74%) for FPV PAT detection, but was highly accurate in the assessment of FHV and FCV PAT. Improvements in accuracy and repeatability of FPV PAT determination could make this tool a valuable component of a disease outbreak response in animal shelters.     

Results of a web-based health survey of retired racing Greyhounds

BACKGROUND: Adoption of retired racing Greyhounds has become increasingly popular during the past decade. To date, research has focused on the physiologic and clinicopathologic peculiarities of Greyhounds but there is little published information on disease prevalence in the breed. OBJECTIVE: The objective of this study is to determine the prevalence of disease in retired racing Greyhounds. ANIMALS: In this study, 747 Greyhounds were used. METHODS: A standardized survey method was used, and survey responses were collected by an Internet survey. Owners could answer a survey for every Greyhound that they had owned since January 1, 2005. RESULTS: Of the 692 eligible participants, 441 (63.7% response rate) completed surveys for 747 Greyhounds. The mortality rate for Greyhounds within the 2-year period was 15% (113 of 747 died). The most common cause of death reported was cancer (66 dogs, 58%), and the most common type of cancer listed as the cause of death was osteosarcoma (28 dogs, 25%). The most commonly reported groups of diseases or disorders were skeletal (232 dogs, 33%), skin (197 dogs, 28%), digestive (132 dogs, 18%), cancer (94 dogs, 13%), and endocrine (85 dogs, 11.9%). Forty-five percent of Greyhounds diagnosed with cancer and 6% of the overall population had osteosarcoma. CONCLUSIONS AND CLINICAL IMPORTANCE: The results of this study can be used by veterinary researchers to continue to investigate the most common diseases in this population. As more retired racing Greyhounds enter the pet population, the results of this study will help educate veterinarians and owners about the most prevalent diseases in the breed.