Use of electrochemical biosensor and gas chromatography for determination of dichlorvos in wheat

Two methods for the determination of dichlorvos in durum wheat by electrochemical assay and gas chromatography, respectively, have been developed. Dichlorvos, an organophosphorus anticholinesterase pesticide, was extracted from wheat with hexane, and the filtered extract was directly analyzed by gas chromatography with nitrogen-phosphorus flame detection (NPD). Recoveries of dichlorvos from milled wheat spiked at 0.25-1.5 microg/g ranged from 96.5 to 100.9%, and the limit of detection was 0.02 microg/g. The electrochemical assay was based on the detection of choline, the acetylcholinesterase product, via a choline oxidase biosensor. An aliquot of the filtered hexane extract was partitioned with phosphate buffer solution, and the organic layer was evaporated prior to electrochemical analysis. A limit of detection of 0.05 microg/g of dichlorvos was obtained with mean recoveries of 97-103% at spiking levels of 0.25-1.5 microg/g. A good correlation (r = 0.9919) was found between the results obtained with the electrochemical and those obtained with the gas chromatographic methods. The electrochemical method was peer-validated in two laboratories that analyzed 10 blind samples (5 duplicates), including a blank and 4 spiked samples with dichlorvos at levels of 0.25, 0.60, 1.00, and 1.50 microg/g. Within-laboratory repeatability (RSDr) and between-laboratory reproducibility (RSDR) ranged from 5.5 to 7.8% and from 9.9 to 17.6%, respectively.     

Impact of Land Use and Urban Runoff on the Contamination of the Sarno River Basin in Southwestern Italy

A study was conducted to determine the contaminantlevel of the Sarno River basin, a small creek located insouthwestern Italy. Six sampling site locations were selected tocoincide with two previous studies conducted in 1975 and 1985 toprovide a basis for comparison. For each location, twelve sampleswere collected over the period October 1997–September 1998.Results indicated a degradation in river water quality, especiallynear the mouth. This was reflected by a dissolved oxygenconcentration of about 0.8 mg L^-1 near the urban areas ofScafati, Pompei and Castellamare of Stabia. At the same locations,BOD₅ and COD values were two to five times higher than theallowable regulatory maximum levels, respectively. About 4 kmfrom the headwaters, the average oxygen concentration was 4.7 mgL^-1, BOD₅ values were close to the standard limit (37 vs 40 mg L^-1), and COD values were above the standard limits(252 vs 160 mg L^-1). Highest concentrations were detectedduring peak tomato production. During this time, settleable solidswere also elevated, two to six times the regulatory standard.Coliform bacteria densities were found in excess of the standardlimits at all sites. Near the head of the river, mean fecalcoliform (22,571 MPN mL^-1) and streptococci densities (14,214 MPN mL^-1) surpassed the regulatory level of 120 and 20 MPN mL^-1, respectively, reflecting the input from the urbansettings of Sarno and agricultural fields. The heavy metaldissolved concentrations were low at all sampling sites. Most ofthe analyzed contaminants increased when comparend with 1985data. BOD₅ increased 5 to 10 times at sites near the urbansettings of Scafati, Pompei and Castellamare di Stabia, whereasCOD increases were twofold at all sites. A decline of chromiumlevels was observed due to the recent effective treatment ofwaste water from the tanning plants.     

Survival times in dogs with right atrial hemangiosarcoma treated by means of surgical resection with or without adjuvant chemotherapy: 23 cases (1986-2000)

OBJECTIVE: To determine survival times in dogs with right atrial hemangiosarcoma treated by means of pericardectomy and tumor resection, with or without adjuvant chemotherapy, and identify complications associated with treatment. DESIGN: Retrospective study. ANIMALS: 23 dogs. PROCEDURE: Dogs were included only if the diagnosis was confirmed histologically. RESULTS: The most common initial complaints included acute collapse (8 [35%] dogs), anorexia or inappetence (8 [35%]), and lethargy (8 [35%]). The most common physical examination abnormalities included muffled heart sounds (12 [52%] dogs), tachycardia (7 [30%]), and weak pulses (7 [30%]). Postoperative complications developed in 12 (52%) dogs; however, most complications were minor. Twenty (87%) dogs were discharged from the hospital. Survival time was significantly longer in the 8 dogs that received adjuvant chemotherapy (mean, 164 days; median, 175 days) than in the 15 dogs that did not receive chemotherapy (mean, 46 days; median, 42 days). Dogs that received chemotherapy were significantly younger and had significantly lower WBC counts than did dogs that did not receive chemotherapy. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that in dogs with right atrial hemangiosarcoma, surgical resection of the tumor was associated with a low complication rate and complications that did arise typically were minor. In addition, use of adjuvant chemotherapy following resection was associated with significantly longer survival times, compared with resection alone.     

Campylobacter

Species within the genus, Campylobacter, have emerged over the last three decades as significant clinical pathogens, particularly of human public health concern, where the majority of acute bacterial enteritis in the Western world is due to these organisms. Of particular concern are the species, C. jejuni and C. coli, which are responsible for most of these gastrointestinal-related infections. Although these organisms have already emerged as causative agents of zoonoses, several aspects of their epidemiology and pathophysiology are only beginning to emerge. Trends in increasing antibiotic resistance are beginning to emerge with oral antibiotics, which may be the drug of choice for when it is necessary to intervene chemotherapeutically. This review wishes to examine (i) emerging clinical aspects of the disease, such as Guillain Barre syndrome (GBS), (ii) the association between these organisms and poultry as a natural host, (iii) environmental aspects of Campylobacter epidemiology, (iv) the emergence of atypical campylobacters (v) emerging trends in antibiotic resistance, (vi) adoption of modern methods for the detection of campylobacters.     

Isolation of Malassezia species from healthy cats and cats with otitis

Lipid-dependent Malassezia species have recently been cultured from veterinary specimens. The identification of Malassezia species isolates from animals is important to clarify the epidemiology of these lipophilic yeasts. Malassezia species were cultured from the external ear canals of 63 out of 99 cats with otitis and 12 of 52 (23%) healthy control cats. The rate of isolation in affected animals versus controls was highly significant (P<0.01). Malassezia pachydermatis was isolated as a pure culture in 33 (45.2%) cats, associated with Malassezia globosa and Malassezia furfur in 20 (50%) and 17 (42.5%) animals, respectively. Three different species were isolated simultaneously in three cats (two cats with M pachydermatis, M globosa and M furfur, one subject with M pachydermatis, M furfur and Malassezia sympodialis). M globosa was isolated as the sole species in two animals. The present work confirms the presence of some lipid-dependent species of Malassezia in both healthy and otitic cats.     

How to obtain a blood pressure measurement

Noninvasive blood pressure measurement is now a standard diagnostic test in small animal practice. Factors such as patient selection, environment, technical support staff, and necessary equipment and supplies will be discussed. Step by step techniques for obtaining reliable blood pressure measurements in both cats and dogs are presented.     

Parasitic health of olive baboons in Bwindi Impenetrable National Park, Uganda

This study examined the endoparasite load of a group of olive baboons (Papio cynocephalus anubis) that share their habitat with a population of mountain gorillas (Gorilla gorilla berengei) in Bwindi Impenetrable National Park, Uganda. During a 2-week period in June 2002, shared habitat was substantiated by noting the ranging patterns of both species and recording observations and trail remains with a global positioning system (GPS). Parasite load was determined by collecting fecal samples preserved in 10% formalin and analyzed by sedimentation techniques and immunofluorescent antibodies (IFA). Bwindi baboons were infected with Cryptosporidium, Giardia, Eimeria, Ascaris spp., Strongyloides spp., Trichuris spp., Hymenolepis spp., and Ternidens diminuta. Of these, the Bwindi mountain gorillas had previously been found to carry Cryptosporidium, Giardia, Strongyloides spp., and Trichuris spp., but not Eimeria, Ascaris, Hymenolepis, or T. diminuta. This study is the first to record Giardia in olive baboons and demonstrates that at least three parasite species in Bwindi baboons have yet to be found in Bwindi gorillas. Early monitoring is essential to the health and well being of both of these sympatric primate species, especially as they continue to interact more often and over a larger area.     

Presumptive fenbendazole toxicosis in North American porcupines

CASE DESCRIPTION: 4 North American porcupines were evaluated because of diarrhea or neutropenia (or both) that developed after treatment with fenbendazole for intestinal parasites. CLINICAL FINDINGS: Complete blood cell count abnormalities included severe neutropenia in all affected porcupines and mild anemia in some of them. In 2 porcupines, postmortem findings included bone marrow hypoplasia and intestinal crypt cell necrosis. TREATMENT AND OUTCOME: Affected porcupines received supportive care including fluid supplementation and broad-spectrum antimicrobials. The 2 surviving animals recovered after 9 to 33 days of treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Fenbendazole is an anthelminthic that may be used in an extralabel manner for the treatment of intestinal parasitism in wildlife species. The drug inhibits mitosis and can affect rapidly dividing cell lines, such as those in the bone marrow and intestinal crypt mucosa. Fenbendazole may not be an appropriate anthelminthic choice in North American porcupines.     

Structure of an intermediate state in protein folding and aggregation

Protein-folding intermediates have been implicated in amyloid fibril formation involved in neurodegenerative disorders. However, the structural mechanisms by which intermediates initiate fibrillar aggregation have remained largely elusive. To gain insight, we used relaxation dispersion nuclear magnetic resonance spectroscopy to determine the structure of a low-populated, on-pathway folding intermediate of the A39V/N53P/V55L (A, Ala; V, Val; N, Asn; P, Pro; L, Leu) Fyn SH3 domain. The carboxyl terminus remains disordered in this intermediate, thereby exposing the aggregation-prone amino-terminal β strand. Accordingly, mutants lacking the carboxyl terminus and thus mimicking the intermediate fail to safeguard the folding route and spontaneously form fibrillar aggregates. The structure provides a detailed characterization of the non-native interactions stabilizing an aggregation-prone intermediate under native conditions and insight into how such an intermediate can derail folding and initiate fibrillation.     

Efficacy of QCDCR formulated CpG ODN 2007 in Nile tilapia against Streptococcus iniae and identification of upregulated genes

The potential of using a QCDCR (quilA:cholesterol:dimethyl dioctadecyl ammonium bromide:carbopol:R1005 glycolipid) formulated CpG oligodeoxynucleotide (ODN), ODN 2007, to confer protection in Nile tilapia against Streptococcus iniae infection was evaluated in this study. At two days post treatment, QCDCR formulated ODN 2007 elicited significant (P<0.05) protection to Nile tilapia, with relative percent survival of 63% compared to fish treated by QCDCR alone. To understand the molecular mechanisms involved in the protective immunity elicited by ODN 2007, suppression subtractive cDNA hybridization technique was used to identify upregulated genes induced by ODN 2007. A total of 69 expressed sequence tags (ESTs) were identified from the subtractive cDNA library. Quantitative PCR revealed that 44 ESTs were significantly (P<0.05) upregulated by ODN 2007, including 29 highly (>10-fold) and 15 moderately (<10-fold) upregulated ESTs. Of all ESTs, putative peroxisomal sarcosine oxidase was upregulated the highest. The 69 ESTs only included six genes that had putative functions related to immunity, of which only two (putative glutaredoxin-1 and carboxypeptidase N catalytic chain) were confirmed to be significantly upregulated. Our results suggest that the protection elicited by ODN 2007 is mainly through innate immune responses directly or indirectly related to immunity.