The bactericidal activity of a teat dip containing chlorhexidine and cetrimide.

Using an in vivo test on teat skin the disinfectant activity of a teat dip containing chlorhexidine and cetrimide was compared with two iodophor solutions, one containing the recommended concentration of 0.5 per cent available iodine and the other a 10-fold dilution of this (0.05 per cent iodine). The test organisms used were Staphylococcus aureus and Escherichia coli and for both species the 0.5 per cent iodophor was significantly more bactericidal than either the diluted iodophor or the chlorhexidine/cetrimide teat dip (P less than 0.01). In the test against S aureus, chlorhexidine/cetrimide and the 0.05 per cent iodophor showed similar bactericidal activity, but the iodophor was significantly more bactericidal against E coli (P less than 0.01). It is argued that due to its low bactericidal activity this formulation of chlorhexidine/cetrimide is likely to be inferior to 0.5 per cent iodophor solution as a disinfectant teat dip.     

The clinical effectiveness of a post milking teat disinfection method with a foaming iodophor teat dip

The effect of postmilking teat dipping with a foaming iodophor agent on incidence of intramammary infections (IMI), incidence of clinical mastitis, somatic cell count and the characteristics of udder tissue and teat was investigated in a positively controlled field study. Two groups of animals were compared. Teats were dipped with a foaming iodophor in the treatment group (TG, 122 animals) while teats in the control group (CG, 121 animals) were dipped with a conventional iodophor teat dip with the same iodine content. A bacteriological examination of quarter milk samples divided the study period in two parts. The incidence of new IMI did not differ between the groups (1st part of trial: TG vs. CG: 6.84% vs. 9.16%, 2nd part of trial: 7.78% vs. 7.82%). There were no differences between the treatment groups regarding incidence of clinical mastitis. We detected 0.64 clinical cases per 100 days in the treatment group vs. 0.50 in the control group. The development of SCC was comparable in both groups. Teat skin and teat duct conditions showed variation during the study period. Clinical efficacy of postmilking teat disinfection with a foaming iodophor was comparable to the treatment with a conventional iodophor product.     

Field evaluation of a teat dip containing dodecyl benzene sulfonic Acid in preventing new mammary gland infections in a dairy herd

The efficacy of a teat dip containing dodecyl benzene sulfonic acid (1.94%) in reducing the incidence of new Staphylococcus aureus infections was demonstrated in a commercial dairy herd to which a number of uninfected cows and heifers were introduced. In the first three months the occurrence of new quarter infections in the teatdipped cows was reduced by 56.49% as compared to the undipped controls.     

Efficacy of a germicidal post milking barrier-type teat dip in preventing intramammary infections

Five commercial dairy herds with 269 lactating cows participated for 12 months in a field trial to determine the effectiveness of a barrier teat dip containing chlorous acid/chlorine dioxide germicide. The right quarters of the cows in two herds and the left quarters of the cows in three herds were dipped in the experimental teat dip after the removal of the milking machine. The other quarters were dipped in a conventional 0.5 per cent iodophor product. Compared with this control product, teat dipping with the experimental dip reduced the number of new intramammary infections by 18.8 per cent, infections with major pathogens by 13.6 per cent, infections with minor pathogens by 16.8 per cent and clinical mastitis by 33.3 per cent. Statistical analysis indicated a trend towards decreased intramammary infections with coliforms and coagulase-negative staphylococci when the experimental teat dip was used. It was concluded that under the conditions of this investigation it was effective in preventing new infections due to both contagious and some environmental pathogens. However, the data suggested that it could adversely affect the condition of the skin of the teat when it was used after incorrect preparation of the udder.     

Prevention of intramammary infections in dairy cows by the use of a premilking teat dip method with a foaming iodophor dip agent

In this study we investigated the efficacy of premilking teat dipping with a foaming iodophor teat dip in a negative controlled field study. Incidence of new intramammary infections (IMI), incidence of clinical mastitis, influence on somatic cell count (SCC) and the characteristics of udder tissue and teats were used as parameters to evaluate clinical efficacy. Predipping was compared with a negative control using a split-udder experimental design. Right teats were predipped with a foaming disinfectant containing 0.27% iodine while left teats served as controls. The latter were conventionally cleaned with damp cloth towels and dried manually with disposable paper towels ("best cleaning practice"). All teats were dipped after milking with the same dip. There were no differences between treated and control quarters with respect to incidence of new IMI during the study period (treated quarters: 6.6% vs. untreated: 6.95%), incidence of clinical mastitis (30 cases in the treatment group vs. 39 cases in the control group) and geometric mean of SCC of quarter milk samples. Spectrum of detected pathogens was also comparable. Condition of udder tissue and teat ducts did not differ between treated and control quarters.     

In vitro and in vivo evaluation of hypericin for photodynamic therapy of equine sarcoids

The therapeutic potential of the photodynamic compound, hypericin, in the treatment of equine sarcoids was evaluated. The in vitro cytotoxicity was assessed using three equine cell lines and the observed phototoxic effect was comparable to that on different highly sensitive human cell lines and significantly influenced by the energy density used although independent of the cell type. The in vivo antitumoural action of photodynamic therapy using hypericin was evaluated on three equine sarcoids in a donkey. Four intratumoural injections were given and the tumours were illuminated daily during 25 days. An 81% reduction in tumour volume was obtained at the end of therapy and 2 months later, a 90% reduction was observed. Further experimental work should be performed, but these results suggest that photodynamic therapy using hypericin has a potential for the non-invasive treatment of equine sarcoids.     

In vitro and in vivo acaricidal activity of a herbal extract

The anti-tick efficacy of combined aqueous herbal extracts of Azadirachta indica leaves, Nicotiana tabacum leaves, Calotropis procera flowers and Trachyspermum ammi seeds was evaluated using adult immersion test, larval packet test and ear bag method. The extract exhibited lethal effects on egg laying (index of egg laying=0.371404±0.00435), hatching (22.35%) and total larval mortality at 50 mg ml(-1) and reduced tick intensity on the infested calves (18 detached out of 35 at 45% (w/w) suspension, topically applied). The herbal extract exerted dose- and time-dependent response against all the developmental stages of Rhipicephalus (Boophilus) microplus considered in this study, thus justified their use in the traditional system of Pakistan.     

The irritancy of chlorhexidine gluconate in the genital tract of the mare

Uterine irrigation was carried out in eight Welsh pony mares using 50 ml of chlorhexidine gluconate solution diluted to give active ingredient concentrations of 0.25 per cent to 2 per cent. Treatment was repeated up to twice in mares showing no adverse effects and irritancy of treatment judged on clinical symptoms and uterine biopsy. Results indicated the inadvisability of using a higher concentration than 0.25 per cent. Three daily applications of a diluted surgical scub solution containing 2 per cent chlorhexidine gluconate to the penis of an arab stallion failed to produce symptoms or irritation. An in vitro minimum inhibitory concentration test performed with the contagious equine metritis organism confirmed its high sensitivity to both chlorhexidine gluconate preparations.     

In vitro antimicrobial activity of a commercial ear antiseptic containing chlorhexidine and Tris–EDTA

Minimum bactericidal concentrations (MBCs) of a commercial ear antiseptic containing chlorhexidine 0.15% and Tris–EDTA (Otodine^®) were determined by broth microdilution for 150 isolates representing the most common pathogens associated with canine otitis. The microorganisms were classified into three groups according to their levels of susceptibility. The most susceptible group included Staphylococcus pseudintermedius, Malassezia pachydermatis, Streptococcus canis and Corynebacterium auriscanis, which were generally killed by 1 : 64 dilution of the antiseptic product (MBC = 23/0.8 μg/mL of chlorhexidine/Tris–EDTA). The most resistant organism was Proteus mirabilis, which survived up to 1 : 8 dilution of the product (MBC = 375/12 μg/mL). Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus displayed intermediate MBCs ranging between 188/6 and 47/1.5 μg/mL. Interestingly, S. pseudintermedius was more susceptible than S. aureus, and no significant difference was observed between meticillin‐resistant and meticillin‐susceptible isolates within each species, indicating that antiseptic use is unlikely to co‐select for meticillin resistance. Although the concentrations required for killing (MBCs) varied considerably with microorganism type, the combination of chlorhexidine 0.15% and Tris–EDTA was active against all the pathogens most commonly involved in canine otitis.     

Effect of teat dipping with a germicide barrier teat dip in late gestation on intramammary infection and clinical mastitis during the first 5 days post-partum in primiparous cows

The effect of teat dipping with a barrier teat dip prior to parturition on intramammary infection (IMI) and clinical mastitis during the first 5 days post-partum was investigated in a split udder trial in 149 Holstein-Frisian heifers. Their left front and right hind quarters were dipped three times weekly (i.e. Monday, Wednesday and Friday) with a barrier teat dip containing 0.1% polyvidon iodine from day 260 of gestation until parturition. The opposite quarters (right front and left hind quarter) served as untreated control. Bacteria were isolated from 52.2% of quarter milk samples collected immediately after parturition prior to first machine milking. Staphylococcus aureus and coagulase-negative staphylococci (CNS) were predominantly found in the samples (29.2 and 35.6% of the positive samples, respectively). At parturition 6.7% of the heifers showed signs of clinical mastitis and another 27.5% developed signs of clinical mastitis during the first five days of lactation. No significant differences were found between treated and control quarters regarding IMI and incidence of clinical mastitis. Teat dipping prior to parturition in primigravid dairy heifers did not improve udder health in this trial.     

In vitro and in vivo evaluation of lysozyme extracts of virulent Mycobacterium bovis in guinea pigs and calves

Extracts of Mycobacterium bovis ATCC 19210 were prepared from cells following treatment with acetone 3 times, ethyl alcohol-ether 3 times (1:1, v/v), and chloroform 3 times. Cells were dried and suspended in 0.05M Tris-HCl (pH 7.5) containing lysozyme (1 mg/50 mg of dried cells). One aliquot of the lysozyme extract was filter-sterilized and 1 aliquot of the lysozyme extract was autoclaved. Delayed-type hypersensitivity responses elicited in sensitized guinea pigs, using the filter-sterilized lysozyme extract, were significantly greater than responses elicited using the autoclaved lysozyme extract (P less than 0.01). The filter-sterilized lysozyme extract and a purified protein derivative (PPD) of M bovis, at equal protein concentrations, elicited comparable delayed-type hypersensitivity responses in sensitized guinea pigs. Significant differences were not detected between the mean enzyme-linked immunosorbent assay (ELISA) values on sera collected from calves before exposure to M bovis, using each of the lysozyme extracts or the M bovis PPD (P greater than 0.05). Significant differences were detected when ELISA values obtained using each of the antigens on post-exposure serum were compared with ELISA values on serum collected from calves before exposure to M bovis (P less than 0.01). Differences were not detected in mean ELISA values on sera from cattle collected 12 months after exposure to M bovis, using each of the lysozyme extracts or M bovis PPD (P greater than 0.05); however, 8 of 8 calves were identified as positive on ELISA, using the filter-sterilized lysozyme extract, 7 of 8 calves were positive, using M bovis PPD, and 7 of 8 calves were positive, using the autoclaved lysozyme extract.     

In vitro growth inhibition of mastitis pathogens by bovine teat skin normal flora.

One factor contributing to differences in the susceptibility of cows to mastitis may be differences in the teat skin normal flora, which could inhibit or enhance the growth of pathogenic bacteria. Using in vitro cross-streaking methods, we found that 25% of the isolates of teat normal flora of non-lactating heifers inhibited the growth of selected mastitis pathogens, but enhancers were not detected. Gram-positive pathogens were inhibited to a greater extent than Gram-negative pathogens. Inhibition was not a characteristic of specific genera or species of normal flora, but rather a property of certain variants within a species. This phenomenon of inhibition of mastitis pathogens in vitro by normal flora may be useful as an in vivo biological control method to reduce the incidence of mastitis.     

An acute outbreak of teat lesions affecting 38% of a dairy herd in Northland

Extract

Teat lesions are common in dairy cattle in New Zealand. Most of the infectious cases are caused by pseudocowpox virus, but lesions similar to those caused by bovine herpesvirus (BHV)-2 have been described clinically and confirmed serologically in New Zealand (Horner and Raynel 1988). BHV-2 and BHV-4 cause an acute viral disease in cattle known as bovine herpes mammillitis or ulcerative mammillitis (Hillerton et al 2001). Presumptive cases have been reported in New Zealand based on the nature and distribution of lesions (Daniel 1970). The disease is probably endemic in New Zealand but is poorly documented and generally considered to be mild and sporadic. The main differential diagnosis is pseudocowpox, which is often also present in herds affected with bovine herpes mammillitis, and dual infections of individual cattle have been reported (Gibbs et al 1972). To my knowledge, neither BHV-2 nor BHV-4 have yet been isolated in New Zealand.     

In vitro and in vivo evaluation of an in situ forming gel system for sustained delivery of Florfenicol

The objective of this study was to develop an injectable in situ forming gel system based on Poloxamer for sustained release of Florfenicol (FFC). The formulations were prepared containing certain amounts of Poloxamer 407 (P407) and Poloxamer 188 (P188) alone or with hydroxylpropyl methylcellulose (HPMC), sodium carboxymethyl cellulose (CMC‐Na), or polyvinyl pyrrolidone (PVP) as polymer additives. The optimal formulation was chosen according to in vitro parameters (gelation temperature, gelation time, pH value, viscosity, and in vitro release). Then the FFC in vivo pharmacokinetic character of the optimal formulation was investigated in dogs with a single dose of 50 mg/kg b.w. under s.c. injection. In vitro release studies, all formulations containing polymer additives had prolonged release time and decreased initial burst to some extent. The optimal formulation containing 0.15% HPMC showed a best sustained release profile for about 128 h with the lowest initial burst in vitro (<40% in 24 h). In vivo, the 20% FFC in situ forming gel provided prolonged drug release time within the therapeutic range for about 100 h, with stable plasma levels and elimination half‐life (t_1/2λz) nine times higher than the control formulation. In conclusion, in situ forming gel is an attractive alternative for FFC sustained release system.