Comparison of a quantitative microtiter method, a quantitative automated method, and the plate-count method for determining microbial complement resistance.

A quantitative microtiter method for determining the degree of complement resistance or sensitivity of microorganisms is described. The microtiter method is compared with a quantitative automated system and the standard plate-count technique. Data were accumulated from 30 avian Escherichia coli isolates incubated at 35 C with either chicken plasma or heat-inactivated chicken plasma. Analysis of data generated by the automated system and plate-count techniques resulted in a classification of the microorganisms into three groups: those sensitive to the action of complement; those of intermediate sensitivity to the action of complement; and those resistant to the action of complement. Although the three methods studied did not agree absolutely, there were statistically significant correlations among them.     

THE POTENTIAL OF SOME SYNTHETIC PYRETHROIDS FOR CONTROL OF THE CATTLE TICK (BOOPHILUS MICROPLUS)

As part of a continuing programme to aid in the development of new acaricides, the potentials of three synthetic pyrethroids, permethrin, cypermethrin and decamethrin, were assessed against a range of resistant strains of the cattle tick (Boophilus microplus). The compounds were shown to be acaricidal, with decamethrin the most effective, giving efficient control of susceptible and organo-phosphorus resistant strains of the cattle tick at concentrations as low as one fifth of those previously used for any commercial acaricide. However higher concentrations were required for satisfactory control of the DDT-resistant strain. Permethrin-selection of a field strain containing a low percentage of DDT-resistant ticks led to rapid development of resistance to a level equivalent to that of the DDT-resistant strain. Further selection increased resistance to permethrin to a higher level than that shown in the DDT-resistant strain. This increased resistance to permethrin appears to be restricted mainly to this compound and is not associated with a marked increase in resistance to cypermethrin, decamethrin or DDT. The potential of organo-phosphorus compounds to potentiate the pyrethroids when the two classes of compounds are used as mixtures against the pyrethroid-resistant and susceptible strains was confirmed.     

Plasma pharmacokinetics and faecal excretion of ivermectin, doramectin and moxidectin following oral administration in horses

The present study was carried out to investigate whether the pharmacokinetics of avermectins or a milbemycin could explain their known or predicted efficacy in the horse. The avermectins, ivermectin (IVM) and doramectin (DRM), and the milbemycin, moxidectin (MXD), were each administered orally to horses at 200 microg/kg bwt. Blood and faecal samples were collected at predetermined times over 80 days (197 days for MXD) and 30 days, respectively, and plasma pharmacokinetics and faecal excretion determined. Maximum plasma concentrations (Cmax) (IVM: 21.4 ng/ml; DRM: 21.3 ng/ml; MXD: 30.1 ng/ml) were obtained at (tmax) 7.9 h (IVM), 8 h (DRM) and 7.9 h (MXD). The area under the concentration time curve (AUC) of MXD (92.8 ng x day/ml) was significantly larger than that of IVM (46.1 ng x day/ml) but not of DRM (53.3 ng x day/ml) and mean residence time of MXD (17.5 days) was significantly longer than that of either avermectin, while that of DRM (3 days) was significantly longer than that of IVM (2:3 days). The highest (dry weight) faecal concentrations (IVM: 19.5 microg/g; DRM: 20.5 microg/g; MXD: 16.6 microg/g) were detected at 24 h for all molecules and each compound was detected (> or = 0.05 microg/g) in faeces between 8 h and 8 days following administration. The avermectins and milbemycin with longer residence times may have extended prophylactic activity in horses and may be more effective against emerging and maturing cyathostomes during therapy. This will be dependent upon the relative potency of the drugs and should be confirmed in efficacy studies.     

Analgesia of newborn lambs before castration and tail docking with rubber rings

This study assessed the effects of suckled sucrose and parenteral carprofen treatment on behavioural measurements of discomfort and serum haptoglobin concentration in lambs following rubber ring castration and/or tail docking. Twenty-eight male and 20 ewe lambs were allocated to either a male or ewe lamb control group (n=4) or to one of three male and two ewe lamb treatment groups (n=8). Male lambs in one treatment group received carprofen subcutaneously (0.5 mg/kg) 30 minutes before castration and docking. Lambs in all other treatment groups suckled sucrose or colostrum immediately before castration and/or tail docking. Behavioural measurements of discomfort were made following castration or tail docking in treatment groups and also in control animals which were not castrated or tail docked. Blood sampling of animals in treatment groups for analysis of serum haptoglobin, an acute phase protein used as an indicator of an acute inflammatory response, was performed before castration or docking and at 24 and 48 hours after castration or docking. Control lambs were blood sampled at 0, 24 and 48 hours following behavioural assessment Neither suckled sucrose nor carprofen treatment altered discomfort behaviour associated with castration or tail docking. Haptoglobin levels following castration or tail docking remained close to the detection limits of the assay and were similar to those recorded in control animals.     

The life cycle of Paracardicoloides yamagutii Martin, 1974 (Digenea: Sanguinicolidae)

The sanguinicolids Paracardicoloides yamagutii Martin, 1974 and Plethorchis acanthus Martin, 1975 were obtained from their definitive hosts, Anguilla reinhardtii Steindachner and Mugil cephalus Linnaeus (respectively) in the tributaries of the Brisbane River, Queensland, Australia. Two putative sanguinicolid cercariae were collected from a hydrobiid gastropod, Posticobia brazieri Smith, in the same waters. The two cercariae differ markedly in size and the form of their sporocysts. Both putative cercariae develop in the digestive gland of Po. brazieri. The ITS2 rDNA region from these sanguinicolids and a Clinostomum species (utilised as an outgroup due to the close morphological similarities between the cercarial stages of the Clinostomidae and the Sanguinicolidae) were sequenced and aligned. Comparison of the ITS2 sequences showed one cercaria to be that of P. yamagutii. This is the first sanguinicolid life history determined by a molecular method. P. yamagutii is the fourth sanguinicolid known to utilise a freshwater hydrobiid gastropod as its intermediate host. ITS2 rDNA is effective in distinguishing sanguinicolids at the species level.     

Maintenance of a captive flock of house finches free of infection by Mycoplasma gallisepticum

Since the beginning of an epidemic of conjunctivitis in wild house finches caused by Mycoplasma gallisepticum (MG), all captive colonies established by capturing free-ranging house finches from the eastern population have also either been infected at the time of capture or developed infection shortly after capture. In an attempt to avoid this infection in captive flocks being maintained for studies of the finches' behavior and ecology, we compared two different flock management strategies and were able to prevent the development of mycoplasmal conjunctivitis with one of the strategies. Single-sex flocks were built by introducing only seronegative wild-caught birds showing no clinical signs of conjunctivitis and covering their outdoor flight cages with netting to prevent interaction with other wild birds although only the female flocks were initially treated with a 6-wk course of tylosin tartrate (0.3 mg/ml). The female flocks never developed conjunctivitis although the disease did develop in the male flocks. Furthermore, serologic assessments of the healthy flock by serum plate agglutination assays for MG indicated that the females remained free of MG infection in the final 7 wk of the study, during which they were unmedicated. We conclude that any low-level MG infection not diagnosed by the initial test for seroconversion was cleared by the prolonged drug treatment.     

Cloning and sequencing of the iss gene from a virulent avian Escherichia coli

Control of colibacillosis is important to the poultry industry. We have found that the presence of a gene for increased serum survival, iss, is strongly correlated with Escherichia coli isolated from birds with colibacillosis. Therefore, the iss gene and its protein product, Iss, are potential targets for detection and control of avian colibacillosis. The iss gene was amplified from a virulent avian E. coli isolate and sequenced. The sequences of the gene and the predicted protein product were compared with those of iss from a human E. coli isolate and lambda bor. The iss gene from the avian E. coli isolate has 96.8% identity with the iss gene from the human E. coli isolate and 89.4% identity with lambda bor. The Iss protein from the avian isolate has 87% identity with Iss from the human isolate and 90% identity with Bor. The low identity between the two Iss proteins is because of a frame-shift in their respective coding sequences. In sum, iss from this avian E. coli isolate is very similar to iss from a human E. coli isolate, but because of a frameshift mutation in the coding sequence of iss from the human E. coli isolate, Iss proteins from avian and human E. coli isolates have only 87% identity. The strong association of iss with E. coli isolated from birds with colibacillosis, suggests that this sequence be studied for its value as a marker or target to be used in colibacillosis control.     

Comparative study of propofol or propofol and ketamine for the induction of anaesthesia in dogs

The effects of propofol alone or propofol and ketamine for the induction of anaesthesia in dogs were compared. Thirty healthy dogs were premedicated with acepromazine and pethidine, then randomly allocated to either treatment. Anaesthesia was induced with propofol (4 mg/kg bodyweight intravenously) (group 1), or propofol and ketamine (2 mg/kg bodyweight of each intravenously) (group 2). Anaesthesia was maintained with halothane, delivered in a mixture of oxygen and nitrous oxide (1:2) via a non-rebreathing Bain circuit. Various cardiorespiratory parameters were monitored at two, five, 10, 15, 20, 25 and 30 minutes after induction, and the animals were observed during anaesthesia and recovery, and any adverse effects were recorded. During anaesthesia, the heart rate, but not the systolic arterial pressure, was consistently higher in group 2 (range 95 to 102 beats per minute) than in group 1 (range 73 to 90 beats per minute). Post-induction apnoea was more common in group 2 (11 of 15) than in group 1 (six of 15). Muscle twitching was observed in three dogs in each group. Recovery times were similar in both groups. Propofol followed by ketamine was comparable with propofol alone for the induction of anaesthesia in healthy dogs.     

Effects of infection with the ectoparasite Argulus japonicus (Thiele) and administration of cortisol on cellular proliferation and apoptosis in the epidermis of common carp,Cyprinus carpio L., skin

The host‐parasite interaction between juvenile carp, Cyprinus carpio, and the ectoparasitic branchiuran, Argulus japonicus, together with the role of cortisol in this interaction, was examined at the level of the host skin epidermis. Epidermal mucous cell numbers, and proliferation and apoptosis of the epithelial cells were studied over 32 days. Apoptotic cell numbers in the uppermost epidermis were reduced at 26 days post‐infection with A. japonicus, while the other parameters were unaffected. Administration of cortisol‐containing food resulted in reduced apoptosis in the cells in the upper skin epidermis at 24 h and at 28 days post‐feeding. Cortisol feeding combined with A. japonicus infection reduced numbers of apoptotic cells in the upper epidermis more than either individual treatment. Further, combining the treatments also significantly increased apoptosis in the lower epidermis in cells morphologically identified as leucocytes apparently migrating macrophages and lymphocytes. Using immunohistochemistry, we demonstrated cortisol receptor presence and cellular localization in the teleost epidermis. Receptors only occurred in pavement cells in the upper epidermis and in leucocytes in the lower parts of the epidermis. The ectoparasites, or administered cortisol, induced effects which may be functionally adaptive in the upper pavement cells, while combining the two treatments also induced changes indicative of immunosuppression.     

Modelling the effects of capture and sea lice [Lepeophtheirus salmonis (Krøyer)] infestation on the cortisol stress response in trout

Measuring baseline cortisol in wild salmonids is problematic because of the stress induced by most capture methods. The cortisol response to different periods of confinement stress was modelled in a laboratory population of rainbow trout [Oncorhynchus mykiss (Walbaum)]. A quadratic model was used to predict baseline (precapture) cortisol levels in these fish. A similar quadratic response pattern of cortisol was observed in wild sea trout (Salmo trutta L.). These were captured in timed sequences on four dates by seine netting in Clew Bay, west coast of Ireland. The estimated precapture cortisol for these fish indicated higher levels soon after migration to sea and a positive correlation with the level of infestation by Lepeophtheirus salmonis (Krøyer). Curvilinearity of the quadratic model was correlated with levels of lice infestation, indicating an enhanced cortisol response to confinement stress during capture when parasitic abundance was high. Estimated precapture cortisol for individual sea trout was positively correlated with numbers of lice although the linear regression had low predictive power. The method provides, for the first time, a means to estimate baseline cortisol levels in wild salmonids, and addresses one of the most difficult problems in working with wild fish. Keywords: Cortisol, trout, Lepeophtheirus salmonis (Krøyer), capture, quadratic model