Investigations into the effects of concentration and duration of exposure to formalin and malachite green on the survival of the larvae and juveniles of the common carp Cyprinus carpio L. and the sharptooth catfish Clarias gariepinus (Burchell).

Prophylactic dip treatments using formalin and malachite green were applied to 4-day old larvae and 12- and 20-day old juveniles of the European common carp, Cyprinus carpio and the African sharptooth catfish, Clarias gariepinus. Treatments consisted of 100 mg/l malachite green for exposure periods of 10, 30 or 90 s and 200 mg/l formalin, administered for 30, 60 or 90 min. Larvae and juveniles of C. gariepinus could be treated with 100 mg/l malachite green for 10 s, or with 200 mg/l formalin for 30 min, with minimum mortalities. Both chemicals affected the survival of the C. gariepinus juveniles, especially the 90 min exposure to formalin. Juveniles of both species were extremely sensitive to 100 mg/l malachite green concentrations.     

Disinfectant susceptibility testing of avian and swine Campylobacter isolates by a filtration method

The susceptibility testing of disinfectants against Campylobacter jejuni and Campylobacter coli strains from broilers and pigs was investigated. The filtration method European standard EN 1040 was adapted to Campylobacter cultures and validated with reference strains. Two disinfectants were tested: 1% benzalkonium chloride active matter, as quaternary ammonium compound, and 0.63% sodium hypochlorite as chlorine-releasing agent. Both disinfectants were effective against the 34 Campylobacter strains tested after 5 min exposure under in vitro conditions. No link between resistance to disinfectants and antibiotics could be observed.     

Treatment of Cyprinus carpio L. and Clarias gariepinus (Burchell) embryos with formalin and malachite green: effect of concentration and length of treatment on their survival.

Malachite green and formalin were investigated as prophylactic dip treatments for developing embryos of Cyprinus carpio and Clarias gariepinus. Based on the findings, recommendations are made on the respective concentrations to be used and exposure periods to be followed for both formalin and malachite green for both fish species.     

Efficacy of five commercial disinfectants and one anionic surfactant against equine herpesvirus type 1

We investigated the influences of various reaction conditions on equine herpesvirus type 1 (EHV-1) disinfection by 5 commercial disinfectants (3 quaternary ammonium compounds [QACs] and 2 chlorine-based disinfectants) and 1 anionic surfactant. QACs at their highest recommended concentrations had no virucidal effect on EHV-1 with a 10-min reaction time at 0°C or a 1-min reaction time at room temperature. Chlorine-based disinfectants achieved EHV-1 disinfection with a 10-min reaction time at −10°C or a 30-sec reaction time at room temperature. In the presence of 5% fetal bovine serum, QACs (except for benzalkonium chloride) showed more stable virucidal effects than did chlorine-based disinfectants. The virucidal effect of the anionic surfactant was almost equivalent to that of the QACs.     

Susceptibilities to antimicrobials and disinfectants in Salmonella isolates obtained from poultry and swine in Thailand

Salmonella enterica isolates from poultry (n=125) and swine (n=132) in Thailand were investigated for antibiotic resistance, susceptibility to disinfectants (benzalkonium chloride (BKC), chlorhexidine digluconate (CHX), zinc chloride and copper sulfate) and cyclohexane tolerance. Forty-two percent were of multiple resistance to antibiotics. The minimum inhibitory concentrations (MICs) of all antibiotics against isolates from swine were higher than that against the isolates from poultry. There were generally few variations in MICs to all disinfectants, indicating that the isolates had either no or only a limited degree of developed resistance to the disinfectants tested. Only 5 isolates (1.9%) were tolerant to cyclohexane. The proton-dependent efflux systems did not play a major role in the reduced susceptibility to BKC and CHX, since susceptibility was not restored when an efflux inhibitor, carbonyl cyanide m-chlorophenylhydrazone (CCCP) was present. Successive exposure to subinhibitory concentrations of BKC and CHX generated mutants resistant to BKC and CHX. A spontaneous BKC-resistant derivative expressed cross-resistance to antibiotics, chloramphenicol and erythromycin. The mechanism responsible for cross-resistance between BKC and antibiotics was not driven by the proton motif force (PMF).     

Action of disinfectants on canine coronavirus replication in vitro

Canine coronavirus (CCoV) is responsible for enteric disease in pups. Infected dogs generally have a rapid recovery, so the virus is highly contagious and the spread of infection is difficult to control. Chemical disinfectants have been widely used in human disease-control programmes to prevent viral infectious diseases from spreading, but to date, there are no studies in the literature on the sensitivity of CCoV to chemical biocides. The present study investigated the sensitivity of CCoV to disinfectants currently used for prophylaxis in kennel and dog breeding locations. The effects of three agents: alkyl-dimethyl-benzyl-ammonium chloride, benzalkonium chloride and didecyl-dimethyl-ammonium chloride, on the infectivity titre of CCoV in A72 cell lines, were studied at different concentrations. Although they may regard a small number of agents, the findings showed that the sensitivity of CCoV to disinfectants varies and the differences are dose correlated. In general, virus inactivation implies a permanent loss of infectivity which can be evaluated in suspensions and hand disinfection tests.     

Malachite green therapy of proliferative kidney disease in rainbow trout: field trials

The results of a series of experiments to investigate the use of the arylmethane dye, malachite green, for the control of proliferative kidney disease in rainbow trout (Salmo gairdneri Richardson) are described. Under field conditions using the bath application method the dye gave good control of the disease with treatments at 1.6 ppm for 40 minutes repeated at seven, 14 and 21 day intervals. Flush treatments were also successful using the same treatment intervals and beginning with 3.2 ppm malachite green. The problems of toxicity and of standardising the exposure of the fish to the dye with flush treatments are discussed.     

影响福尔马林漂白粉石灰浆消毒效果诸因素的探讨

研究比较了有机氯消毒剂与无机氯消毒剂水溶液的稳定性,药剂液温、药液粘度、pH值对蚕用消毒剂的三种主剂(福尔马林、漂白粉、石灰浆)的不同影响,三种药剂对土壤地面的不同消毒效果以及相互配合使用时会产生的后果。目的在于进一步加深对消毒剂不同性能的认识,为提高消毒效果提供参考。     

Determination of the optimum concentration of decontaminants for the primary isolation of Mycobacterium bovis

The majority of tissue specimens submitted for the isolation of Mycobacterium bovis contain contaminating microorganisms and therefore require selective decontamination before bacteriological examination. The purpose of this study was to identify the preferred decontaminant amongst four commonly used reagents. The four decontaminants used in the study were 1-hexadecylpyridinium chloride, sodium hydroxide, benzalkonium chloride and oxalic acid. A comparison was made of the toxicity of the four decontaminants for M. bovis and their ability to control contamination. Used at the recommended concentrations, all reagents showed a significant degree of toxicity. The toxicity of the decontaminants for three field strains and one laboratory strain of M. bovis were similar, but a second laboratory strain, AN5, was more susceptible. It was also observed that as the concentration of each reagent decreased an abrupt change from control to lack of control of contaminating micro-organisms occurred. Hexadecylpyridinium chloride was found to be the best all-round reagent because at concentrations that effectively controlled contamination this reagent was the least toxic to M. bovis. A protocol for handling specimens based on an assessment of the risk of contamination is recommended.     

Effects of ionic salts or acetylcholine, or both, on electrically induced ventricular fibrillation in dogs

A dose-response curve was delineated for an acetylcholine-potassium chloride cocktail previously found to be highly effective in chemically defibrillating electrical induced ventricular fibrillation of thoracotomized dogs. The cocktail dosage responsible for the greatest percentage of chemical defibrillations was transthoracically given to nonthoracotomized dogs subjected to ventricular fibrillation; this resulted in 33% chemical defibrillation. Two clinical canine patients were transthoracically administered the acetylcholine-potassium chloride cocktail because they failed to respond to external electrical defibrillation; both were chemically defibrillated. Potassium citrate when substituted for potassium chloride, in the cocktail, or alone appeared to be more efficacious for chemical defibrillation. Magnesium chloride with acetylcholine failed to chemically defibrillate any dog administered this cocktail.     

Possible associations between Salmonella persistence in poultry houses and resistance to commonly used disinfectants and a putative role of mar

A putative link between Salmonella persistence in the agricultural sector and resistance to disinfectants has been sparsely investigated. Therefore, minimum inhibitory concentration (MIC) tests against five disinfectants commonly used in poultry premises (formaldehyde, glutaraldehyde/benzalkonium chloride compound, oxidising compound, tar oil phenol, iodophor) were performed on 286 Salmonella isolates, including 256 from Danish broiler houses, altogether representing nine serotypes. Six of these isolates were used for adaptation and de-adaptation studies involving the five disinfectants. Amongst 60 of these isolates selected for growth studies in cyclohexane (possibly associated with up-regulated efflux), only one isolate grew. From this isolate and the six isolates used in the adaptation and de-adaptation studies, mutants highly resistant to triclosan (a disinfectant linked with mar-type resistance) were selected. In addition, adaptation and de-adaptation studies with triclosan were performed. For the 286 isolates, the small variations in MICs could not be associated with Salmonella persistence in Danish broiler houses or previous use of relevant disinfectants. Adaptation and de-adaptation did not alter MICs to the five farm disinfectants. Compared to the parent isolates, MICs for the triclosan adapted and de-adapted isolates and the triclosan mutants were significantly increased to triclosan, but not to the five disinfectants. Moreover, most of the triclosan adapted and de-adapted isolates grew in cyclohexane. Thus, there was no correlation between triclosan and cyclohexane resistance on one hand and resistance to the five disinfectants on the other, suggesting that triclosan resistance is not linked with resistance to these disinfectants.     

Phenotypic Resistance to Disinfectants and Antibiotics in Methicillin‐Resistant Staphylococcus aureus Strains Isolated from Pigs

The aim of this research was to study the phenotypic resistances to disinfectants and antibiotics in strains of methicillin‐resistant Staphylococcus aureus (MRSA) obtained from Canary black pigs. Analyses were performed on 54 strains of MRSA, isolated in Canary black pigs from the province of Tenerife (Spain); all of them carried the mecA gene. The strains were isolated by means of nasal swab samples of healthy pigs, collected under veterinarian supervision. Bactericidal activity of antiseptics and disinfectants was tested by means of the dilution–neutralization method. Susceptibility to the disinfectants glutaraldehyde, peracetic acid and silver nitrate was assessed, as well as to the antiseptics chlorhexidine, benzalkonium chloride and povidone iodine. Susceptibility to a wide array of antibiotics representing the main groups was determined by means of the disc diffusion method. All the strains demonstrated susceptibility to the disinfectants tested at the recommended concentration, and even to dilutions equal to or lesser than 1/16. The most effective antiseptic and disinfectant were, respectively, chlorhexidine and silver nitrate. With regard to the antibiotics, the strains proved to be multiresistant. All presented phenotypic resistance to the β‐lactam antibiotics ampicillin, penicillin and cefoxitin, as well as to numerous aminoglycosides, tetracycline and trimethoprim–sulfamethoxazole. It was also observed that 61.1% of the strains were carriers of plasmids. Our results underline that in the strains such as MRSA, which show multiple resistances to antibiotics, the antiseptics and disinfectants show great efficacy. Moreover, as other authors also suggest, for the treatment and prevention of infections caused by MRSA, the use of β‐lactam and aminoglycoside antibiotics may be less effective.     

The role of preparation technique, culture media and incubation time for embryonation of Heterakis gallinarum eggs

The importance of preparation technique, culture media and incubation time in the embryonation of the infective egg stages of the intestinal nematode parasite Heterakis gallinarum was studied. Mature H. gallinarum worms were isolated from the caeca of infected chickens and separated by sex. In a first experiment intact female worms were kept for the development of their eggs in four different media (0.5% formalin, 2% formalin, 0.1 N sulphuric acid, 0.1% potassium dichromate) and incubated under constant temperature (20-22 degrees C) for 2, 4, 6 or 8 weeks. Afterwards the body of the worms were ruptured and the numbers of unembryonated and embryonated eggs were determined using a McMaster egg counting chamber, and the percentage of embryonated eggs was calculated. After 8 weeks of incubation in 0.5% formalin, 0.1 N sulphuric acid or 0.1% potassium dichromate 27.6%, 26.7% and 29.4% of the eggs, respectively, embryonated into third stage larvae (p > 0.05). In contrast, incubation in 2% formalin resulted in an embryonation of 18.6% only (p < 0.05). In a second experiment H. gallinarum eggs were directly harvested from worm uteri and cultivated afterwards in different media (2% formalin, 0.1 N sulphuric acid, 0.1% potassium dichromate) at 20 to 22 degrees C for 6 weeks. An incubation of isolated eggs in 2.0% formalin or 0.1% potassium dichromate during 6 weeks resulted in a significantly higher percentage of embryonation in comparison to the incubation of intact worms (first experiment). The results suggest that preparation technique, media and time of incubation has an essential influence on the development rate of H. gallinarum eggs.     

Virucidal efficacy of nine commercial disinfectants against porcine circovirus type 2

A number of commercially available disinfectants are commonly used on pig breeding farms and are authorised by the French Agricultural Ministry. However, the efficacy of these disinfectants is unknown with regard to the emergent porcine circovirus type 2 (PCV2). The virucidal efficacy of nine disinfectants was evaluated by testing a suspension of PCV2 isolated in France. The assays were performed at 20 degrees C and the efficacy determined after 30 min contact time between virus and disinfectant. After this time, the mixture was passed through a detoxification column and then diluted to remove compounds toxic to the virus and the porcine kidney cell line. The filtrate was serially diluted and inoculated onto cell culture. The infectivity of PCV2 was determined by an immunoperoxidase monolayer assay. No reduction in PCV2 titre was demonstrated with iodine and phenolic products. Significant PCV2 titre reductions (1.61 log(10)) were noted for the seven other products. For five disinfectants, namely a product composed of potassium monopersulfate, two products comprising a quaternary ammonium with one or three aldehyde(s), sodium hypochlorite, and sodium hydroxide, the concentration that significantly reduced the PCV2 titre was equal or 1.5-4 times lower than the authorised use concentration. Only two disinfectants, one composed of potassium monopersulfate, the other containing peracetic acid with hydrogen peroxide, reduced the PCV2 titre with a product concentration at best equal or two times higher than the authorised use concentration.     

Terminal disinfection of calf houses by formaldehyde fumigation

Trials were carried out to investigate the effectiveness of various methods of formaldehyde fumigation as a means of disinfecting calf houses. Houses were cleaned by the farmer, sealed and then fumigated. A significant reduction in bacterial numbers was obtained when the gas was produced by heating paraformaldehyde, mixing formalin with potassium permanganate or boiling formalin in calf houses that could be effectively sealed. Aerosol generators did not give satisfactory results. Efficient pre-cleaning and sealing of the houses were of paramount importance; relative humidity and temperature were less important.     

Effects of formalin, chloramine-T, and low salinity dip on the behavior and hemolymph biochemistry of the American lobster.

The purpose of this research was to investigate the salinity and formalin sensitivity of a ciliate parasite (Anophryoides haemophila) of the American lobster (Homarus americanus), and to examine the target-animal (lobster) safety of chemical-bath treatments involving low salinity, formalin, or chloramine-T that could be used to control this parasite in lobster pounds. "Bumper car" disease, caused by An. haemophila, is an important concern to lobster pound operators in eastern North America, because of the implicated lobster mortality rate and the general lack of preventive and therapeutic intervention regimes. We determined, using an in vitro method, that formalin at 50 mg/L, or low salinity at 8.0 parts per thousand (ppt) for 1 hour killed 100% of the parasites. When healthy lobsters were exposed to formalin at 200 mg/L, there were no negative behavioral responses and no significant differences in a panel of hemolymph biochemical indices. Similar results occurred when lobsters were exposed to chloramine-T, a common finfish therapeutic agent for topical bacteria and protozoa, at 10 mg/L for 1 hour. The low salinity treatment (8.0 ppt) resulted in significant adverse changes in lobster behavior and biochemical indices; however, these changes did not persist for more than 1 week after treatment ended. Although these treatments are unlikely to kill parasites that have already invaded the lobster carapace, they should be effective in reducing parasite loads on the gill and carapace surface of the lobster and in the environment of the impoundment housing.     

Application of peracetic acid for the treatment of juvenile sander (Sander lucioperca) during Ichthyophthirius multifiliis infestation

White spot disease caused by the ciliate protozoan parasite Ichthyophthirius multifiliis (I. m.), is one of the most dangerous diseases in aquaculture and ornamental fish breeding worldwide. The parasite is characterized by three developmental stages: a reproductive tomont, an infective theront and a parasitic trophont. In sander (Sander lucioperca) breeding I. m. causes serious economic losses. After banning of the traditional therapeutic agent malachite green we have to face a state of emergency for the treatment of the ichthyophthiriasis in Germany. The peracetic acid (PAA), characterized by positive therapeutical properties, might close this gap. The purpose of our investigations was the determination of the toxicity of PES to juvenile sander as well as the evaluation of the therapeutic effectiveness of the substance to combat I. m. For juvenile sander (length 3 cm) we determined a 24-h-LC50 of 1.14 (0.97; 1.3) ppm PES. In two investigations PAA was applied in daily intervals of 0.5; 1, 3, 5 and 24 h and concentrations of 0.5; 1; 1.5 and 2 ppm to treat I.-m.-infection in sander. In test I all sander (length 9 cm) died as a result of the I.-m.-infection. However, the PAA exposed fish survived longer than the PES-free controls! This might be due to a disinfection of other pathogens by PAA. In test II, the fish (length 12 cm) were less infected than fish in test I. Four of six fish died in the group exposed with 2 ppm PAA for 24 hours. The abundance of I.-m.-trophonts was determined in mucus, fin and gill tissues of all fish. Significant differences could not be observed between test I and test II because of dissimilar: 1. exposure in time and concentration, 2. age and condition of the fish and 3. homogenity of the variances. Therefore, no therapy strategy was successful while fighting the parasitic trophonts protected by the overlaying fish tissues. We speculate that a successful therapy of I. m. with PAA is, as known e.g. for malachite green, only possible while fighting the free living stages theronts and tomonts. This will be part of our subsequent investigations.     

Effect of Common Aquaculture Chemicals against Edwardsiella ictaluri and E. tarda

Abstract

Edwardsiellosis is an important bacterial infection of freshwater and marine fishes. Edwardsiella ictaluri causes enteric septicemia of catfish, and E. tarda causes emphysematous putrefactive disease of catfish and fish gangrene in various species; these diseases have considerable economic effects on the aquaculture industry. In addition, E. tarda is an important zoonotic pathogen. Thus, the reduction or elimination of these pathogens from an aquarium or aquaculture facility is imperative. This study examined a variety of commercially available chemicals for their ability to reduce or eliminate E. ictaluri and E. tarda from the aquatic environment. The various concentrations of chemicals were tested in vitro in microcentrifuge tubes with a known concentration of bacteria at room temperature. In this study, ethyl alcohol (30, 50, or 70%), benzyl-4-chlorophenol/phenylphenol (1%), sodium hypochlorite (50, 100, 200, or 50,000 mg/L), n-alkyl dimethyl benzyl ammonium chloride (1:256), povidone iodine (50 or 100 mg/L), glutaraldehyde (2%), and potassium peroxymonosulfate/sodium chloride (1%) were effective disinfectants, as each reduced or eliminated the number of detectable organisms within 1 min of contact time. However, neither Chloramine-T (15 mg/L) nor formalin (250 mg/L) substantially reduced bacterial counts even after 60 min of contact time.     

Bioassay-guided isolation and identification of active compounds from Semen pharbitidis against Dactylogyrus intermedius (Monogenea) in goldfish (Carassius auratus)

The present study was undertaken to isolate the active compounds responsible for the anthelmintic activity of methanol extract of Semen pharbitidis against Dactylogyrus intermedius in goldfish (Carassius auratus). The active methanol extract was fractionated on silica gel column chromatography in a bioassay-guided fractionation, eventually yielding two bioactive compounds: palmitic acid and pharnilatin A by comparing spectral data (NMR and ESI-MS) with literature values. According to in vivo anthelmintic assays, they were found to be 50% effective at the concentrations (EC(50)) of 5.3 and 1.4 mg L(-1), respectively. The promising palmitic acid and pharnilatin A from S. pharbitidis were also subjected to acute toxicity tests for the evaluation of their safety to the host (goldfish). After 48h exposure, the mortalities of goldfish were recorded, and the established LC(50) values were 2.45- and 5.29-fold higher than the corresponding EC(50), demonstrating that pharnilatins A may have better application potential than palmitic acid. The present results provide evidence that pharnilatins A might be potential source of new anti-parasitic drug for the control of Dactylogyrus.     

Hypernatremia secondary to homemade play dough ingestion in dogs: a review of 14 cases from 1998 to 2001

Objective: To correlate the sodium chloride dosage and the serum sodium concentration to clinical signs, to determine if the dosage of homemade play dough (and, therefore, sodium chloride) is the most reliable way to predict clinical signs and prognosis, and to review previously reported treatment options. Design: Retrospective case series. Animals: Fourteen dogs with a history of homemade play dough ingestion. Procedure: Cases were examined for each animal's signalment including body weight, age, approximate amount of play dough ingested, the estimated sodium chloride dosage, clinical signs, serum sodium concentration, and outcome when available. The literature was reviewed to determine pathophysiology and treatment regimens. Results: Twelve of 14 dogs (86%) that ingested homemade play dough showed clinical signs. Vomiting (9 of 14, 64%), polydipsia, and seizures (4 of 14 each, 29%) were the most common signs followed by polyuria, tremors (3 of 14 each, 21%), and hyperthermia (2 of 14, 14%). The lowest calculated dosage associated with objective clinical signs was 1.9 g/kg. Seizures were reported in all animals with serum sodium levels greater than 180 mEq/L. Conclusions and clinical relevance: Homemade play dough ingestion can be a serious and life‐threatening problem. Many factors can contribute to the toxicity of homemade play dough. This study indicates that the serum sodium concentration is a more reliable indicator of the clinical course of the toxicity rather than the amount of play dough and, therefore, the dosage of sodium chloride ingested. Treatment should be based on a clinical evaluation of the patient and laboratory results, and consists of controlling seizures, reducing serum sodium concentrations slowly, and supportive care.