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Biomedical subjects

R B Davey

Publications and source records attributed to R B Davey.

At least 19 recordsLinked to original sources

Molecular cloning and nucleotide sequence of a new P450 gene, CYP319A1, from the cattle tick, Boophilus microplus.

We have isolated and sequenced a novel P450 gene (CYP319A1) from the cattle tick, Boophilus microplus. The CYP319A1 cDNA encodes a protein of 531 amino acids with an estimated molecular weight of 60.9k. It contains all highly conserved motifs characteristic of P450 enzymes. Comparison of deduced amino acid sequence with other CYP members shows that the CYP319A1 is more closely related to CYP4 family, but its overall identity to the CYP4 family is less than 40%. Therefore, it was assigned to a new P450 family by the P450 nomenclature committee. A pseudogene which shares high homology with the CYP319A1 was identified. Analysis of genomic sequence of the pseudogene indicated that the pseudogene contains two additional DNA inserts in the coding region, which disrupt the open reading frame. RT-PCR analysis showed that CYP319A1 is expressed in both susceptible and acaricide-resistant ticks.

Amino Acid Sequence↗

Efficacy of a single whole-body spray treatment of spinosad, against Boophilus microplus (Acari: Ixodidae) on cattle.

The efficacy of a single whole-body spray of spinosad, a naturally derived control agent, applied at three concentrations was evaluated against cattle infested three separate times prior to treatment and at four weekly intervals following treatment with Boophilus microplus (Canestrini). At 0.0167% active ingredient (AI) both tick numbers (1894 ticks per calf) and index of fecundity (IF) of females (258.3) were no different than that of the control group. However, spinosad treatment at both 0.05 and 0.15% AI resulted in fewer ticks per calf (600 and 935, respectively) with lower IF values for females (43.4 and 38.4, respectively). The percent control of ticks on the animals at the time of treatment (acute efficacy) was dramatically lower at 0.0167% AI (21.4%) than at 0.05 (86.3%) and 0.15% AI (87.9%). Spinosad treatments appeared to be more effective against immature stages (nymphs and larvae) than against adult ticks that were on the animals at the time of treatment. The mean weight of females that survived to repletion was similar (322-348 mg) in all groups. By contrast, the mean weight of egg masses produced by females was highest in the control group (155 mg), whereas each increase in spinosad concentration resulted in a substantial decrease in egg mass weight, with the 0.15% AI group averaging only 73 mg. The hatch rate of eggs derived from females ranged from 93.4% in control females down to 53.9% hatch for females treated at 0.15% AI spinosad. The residual efficacy of spinosad at 0.0167% AI was poor even at 1 week following treatment, resulting in 101 ticks per calf and a level of control of only 66.4%. At 0.05% AI, protection against successful reinfestation was high at 1-week post-treatment where only five ticks per calf reached repletion, and control of the IF of these females was 99.3%. The 0.15% AI treatment provided almost complete protection against reinfestation for 2 weeks following treatment (< or =5 ticks per calf), and control of the IF of these ticks was >99.9%. Thus, the use of spinosad at US ports-of-entry would be unacceptable because of the critical necessity of achieving 100% control with a single treatment to prevent the reintroduction of ticks. However, it is likely ticks could be eradicated using spinosad in tick infested areas of the US if repeated (systematic) treatments were applied to cattle maintained on the premises.

Administration, Topical↗

The Ivomec SR Bolus for control of Boophilus annulatus (Acari: Ixodidae) on cattle in South Texas.

When Hereford heifers infested with Boophilus annulatus (Say) were treated with a single Ivomec SR Bolus, the concentration of ivermectin in the serum of the treated cattle reached a maximum of 8.8 +/- 0.9 ppb at 2 wk posttreatment. The single bolus treatment resulted in 84.4% control of standard engorging B. annulatus females on treated cattle over the 20-wk trial. Although fewer engorged ticks were collected from the sentinel heifers exposed in the treated pasture than those in the control pasture at weeks 4, 10, and 16 posttreatment, none of the differences was statistically significant. Each exposure of sentinel cattle found free-living ticks in both the treated and control pastures, indicating the infestation was not eliminated by the treatment. When the trial was repeated using two Ivomec SR Boluses/heifer, the concentration of ivermectin in the serum of the treated cattle reached a maximum level of 31.2 +/- 3.9 ppb at week 13 posttreatment. The use of two boluses/heifer resulted in 99.6% control of standard engorging B. annulatus females over the 20-wk trial. No ticks were found on sentinels placed in the treated pasture after week 9 posttreatment, an indication that the treatment had eliminated the free-living population in the treated pasture. From these studies, we conclude that a single Ivomec SR Bolus is incapable of sufficient control of B. annulatus to meet the rigid requirements of the Cattle Fever Tick Eradication Program in South Texas. Although two boluses per animal did eliminate the ticks from treated heifers and the pasture they were in, the treatment would not be sufficiently efficacious for mature cattle (>400 kg) for it to be useful in the program.

Animals↗

Use of an allele-specific polymerase chain reaction assay to genotype pyrethroid resistant strains of Boophilus microplus (Acari: Ixodidae).

A polymerase chain reaction-based assay was developed to detect the presence of a pyrethroid resistance-associated amino acid substitution in Boophilus microplus (Canestrini). The assay uses a simple method for the extraction of genomic DNA from individual larvae and genotypes individuals for the presence of a Phe-->Ile amino acid substitution in the S6 transmembrane segment of domain III of the para-like sodium channel, clearly distinguishing heterozygotes from homozygotes. High frequencies for this amino acid substitution were found in the Corrales and San Felipe strains, which have target site insensitivity mechanisms for pyrethroid resistance. The Caporal resistant strain contained lower yet substantial numbers of amino acid-substituted alleles. Low amino acid substitution frequencies were found in the susceptible reference Gonzales strain and the Coatzacoalcos strain, which has metabolic esterase-mediated pyrethroid resistance. The amino acid substitution was not found in six other strains that were susceptible to pyrethroids.

Alleles↗

Unusual donor site reactions to calcium alginate dressings.

Calcium alginate dressings have been used as the standard dressing for split skin donor sites at the Women's and Children's Hospital burn unit for the last 12 yr. This method has proven satisfactory with good haemostasis, reliable healing, low infection rate and is well accepted by the children and nursing staff. We have recently had 5 cases with the unusual and unexplained phenomenon of dermal calcification in the donor site following the use of two new varieties of calcium alginate dressing.

Alginates↗

Identification of a point mutation in the para-type sodium channel gene from a pyrethroid-resistant cattle tick.

To investigate the molecular mechanism of resistance to pyrethroids in the southern cattle tick, Boophilus microplus, we have obtained and sequenced a partial para-homologous sodium channel cDNA from susceptible and pyrethroid-resistant tick strains. A point mutation that results in an amino acid change from Phe to Ile was identified in the highly conserved domain IIIS6 of the homologous sodium channel from ticks that are highly resistant to pyrethroid acaricides. This mutation is at a location different from those reported in the same gene in pyrethroid-resistant insects.

Amino Acid Sequence↗

The changing face of burn care: the Adelaide Children's Hospital Burn Unit: 1960-1996.

There have been considerable changes in burn care since the inception of the Adelaide Children's Hospital Burn Unit in 1960, resulting in a marked improvement in both morbidity and mortality. A burn of 30% T.B.S.A. is no longer considered life threatening, whilst a burn of 80% T.B.S.A. is now not only expected to survive, but to have a satisfactory functional and cosmetic outcome. An outline of these changes, as reflected in a review of 37 years of care, is discussed with particular reference to the improvements in scar management.

Adolescent↗

Computerised colour: a technique for the assessment of burn scar hypertrophy. A preliminary report.

A technique for the objective measurement of one parameter of burn scar hypertrophy is described. The technique involves capturing a video camera image on a computer and subsequent quantitative analysis of the colour of the scar using a custom-written computer programme. By analysing multiple areas and comparing the identical areas over the course of treatment, it is theoretically possible to measure the progress and compare differing modes of therapy.

Burns↗

Characterization and molecular cloning of a glutathione S-transferase gene from the tick, Boophilus microplus (Acari: Ixodidae).

A glutathione S-transferase (GST) was purified from the larval cattle tick, Boophilus microplus (Acari: Ixodidae), by glutathione-affinity chromatography. The purified enzyme appeared as a single band on SDS-PAGE and has a molecular mass of 25.8 kDa determined by mass spectrometry. The N-terminus of the purified enzyme was sequenced. The full-length cDNA of the enzyme was isolated by RT-PCR using degenerate oligonucleotides derived from the N-terminal amino acid sequence. The cDNA contains an open reading frame encoding a 223-amino-acid protein with the N-terminus identical to the purified GST. Comparison of the deduced amino acid sequence with GSTs from other species revealed that the enzyme is closely related to the mammalian mu class GST.

Amino Acid Sequence↗

Traumatic pancreatitis: long-term review of initial non-operative management in children.

OBJECTIVE: To assess the safety and long-term efficacy of an initial non-operative approach to the management of pancreatic trauma in children. METHODOLOGY: Retrospective case-note review of children admitted to our institution with the diagnosis of pancreatic trauma over a 10-year period from 1985 to 1994 inclusive. RESULTS: Fourteen children were identified with a diagnosis of traumatic pancreatitis. Eleven of the injuries were secondary to motor vehicle accidents or trauma from a bicycle handlebar. In seven patients there were associated intra-abdominal injuries and in eight patients the traumatic pancreatitis was complicated by development of a pseudocyst. Three pseudocysts settled without intervention, two resolved with percutaneous external drainage and the remaining three required open surgery. The mean in-patient stay was 21.2 days. The average length of follow-up was 7.7 years with no observed long-term complications. CONCLUSIONS: Non-operative management of traumatic pancreatitis in the absence of complete duct transection is safe in children and does not appear to be associated with adverse sequelae.

Accidents, Traffic↗

Control of Boophilus annulatus (Acari: Ixodidae) on cattle using injectable microspheres containing ivermectin.

The efficacy of an injectable microsphere formulation of ivermectin for control of the cattle tick, Boophilus annulatus (Say), was tested on 2 groups of 6 Hereford heifers held on separate 7-ha, tick-infested, buffel grass pastures. Cattle in one pasture were injected subcutaneously in the neck with a controlled-release microsphere formulation of ivermectin at the rate of 2.4 mg AI/kg body weight; the other group was injected with carrier only. Beginning 4 wk after injection and continuing throughout the remainder of the test (16 wk), no engorged ticks (> or = 5.5 mm) were found on any of the treated cattle, whereas large numbers of engorged ticks were found on the untreated controls. During this period, a few ticks were recovered from untreated sentinel animals placed in the treatment pasture during 7-8 wk after treatment, but none were recovered from animals exposed from 11-12 wk or 14-15 wk. Large numbers of B. annulatus ticks were found on untreated sentinel cattle placed in the control pasture during these same periods. Although the cattle, pastures, and tick habitat were approximately equal, the treated cattle gained an average of 77 kg compared with an average of 42 kg for the control group. This technology offers a possible alternative to the current official program of dipping and vacating pastures for eradication of Boophilus sp. infestations from the quarantine zone in southern Texas. Larger scale testing is needed to determine the potential of the injectable microsphere formulation and to optimize its use in eradication or control strategies.

Analysis of Variance↗

Efficacy of coumaphos applied as a dip for control of an organophosphorus-resistant strain of Boophilus microplus (Acari: Ixodidae) on cattle.

The efficacy of coumaphos, an organophosphorus acaricide, was tested on a strain of Boophilus microplus (Canestrini) that was 9.5 times more resistant to the chemical than a susceptible strain. Cattle infested with larval, nymphal, and adult ticks were dipped into a vat charged at concentrations of 0.031, 0.083, 0.160, and 0.279% (AI). Assessment of efficacy was based on surviving tick numbers, female weight, egg mass weight, egg hatchability, and index of reproduction. At 0.031% (AI), the level of control was lower than the other coumaphos treatments, and the weight, fecundity, and fertility of surviving females was similar to untreated females, indicating only minimal adverse effects on the ticks. At 0.160 and 0.279% (AI), no differences in control, female weight, egg mass weight, or percentage egg hatch were observed, but effects were greater than that of untreated ticks or ticks treated at 0.031% (AI). At 0.083% (AI), most biological parameters were intermediate, but there was no difference in control compared with the 2 higher concentrations. Thus, even with the increase in concentration of more than 3 times between treatments of 0.083 and 0.279% (AI), there was no substantial increase in control. The residual efficacy of all 4 treatments showed that none provided complete protection against larval reinfestation for even 1 wk after treatment. Therefore, the presence of organophosphorus-resistant ticks at U.S. ports-of-entry could jeopardize the success of the Boophilus eradication program.

Animals↗

Characterization of pyrethroid resistance and susceptibility to coumaphos in Mexican Boophilus microplus (Acari: Ixodidae).

Two patterns of pyrethroid resistance were characterized from Boophilus microplus (Canestrini) collected in Mexico. One was characteristic of a kdr mutation and the other involved esterase and cytochrome P450 enzyme systems. Very high resistance to permethrin, cypermethrin, and flumethrin, not synergized by TPP and PBO and high resistance to DDT, characterized the kdr-like pattern found in the Corrales and San Felipe strains. Esterase and cytochrome P450-dependent resistance was found in the Coatzacoalcos strain. It was characterized by resistance to permethrin, cypermethrin, and flumethrin, synergized by TPP and PBO, but no resistance to DDT. The Coatzacoalcos strain also showed 3.6-fold resistance to the organophosphate coumaphos. This factor appeared to be independent of pyrethroid resistance. Pyrethroid resistance patterns found in Mexico were similar to those found earlier in Australia. The significance of pyrethroid and coumaphos resistance to the U.S. cattle fever tick quarantine is discussed.

Animals↗

Sequence analysis of the knockdown resistance-homologous region of the para-type sodium channel gene from pyrethroid-resistant Boophilus microplus (Acari: Ixodidae).

Using reverse transcription polymerase chain reaction and degenerate oligonucleotide primers, a partial para-homologous sodium channel cDNA was obtained from the southern cattle tick, Boophilus microplus (Canestrini). The cDNA sequence encoded the region in which knockdown resistance (kdr)-type mutations have been identified in numerous insect species. Comparison of deduced amino acids from the cDNA sequence showed high similarity with sodium channels from other species, particularly in highly conserved repeat domains of the sodium channel. Analysis of the kdr-homologous region of the genomic DNA sequences from several susceptible and pyrethroid-resistant tick strains did not detect mutations. The result suggests novel mutations in the sodium channel gene or metabolic detoxification may be involved in the resistance to pyrethroids in this tick.

Amino Acid Sequence↗

Efficacy of amitraz (Taktic 12.5% EC) as a dip for the control of Boophilus microplus (Canestrini) (Acari: Ixodidae) on cattle.

Four groups of cattle infested with Boophilus microplus (Canestrini) were each dipped in a different concentration of amitraz diluted from a 12.5% EC formulation to determine the efficacy and performance of the product in an 11,400 l dipping vat. Except for the period when heifers were dipped, animals were restrained in stanchions placed individually inside 3.3 x 3.3 m2 stalls within an open-sided barn. The amitraz in the vat was stabilized with hydrated lime to maintain a pH of ca. 12. Analyses of vat samples showed that concentrations of amitraz in the vat were 7.6 to 13% lower than the targeted concentrations of 0.010, 0.015, 0.020, and 0.025% active ingredient (AI) for dilutions prepared according to instructions on the manufacturer's label. The large quantity of hydrated lime added to the vat (10 kg/1000 l) interfered with the HPLC analysis of vat samples. Therapeutic efficacy of each of the four observed concentrations (0.0088, 0.0131, 0.0174, and 0.0231% AI) of amitraz was excellent (> 99% control). However, the rapid detachment of all ticks from an animal within a few hours after treatment with amitraz, that has been frequently observed, was not pronounced in the present study. Only 47% of the B. microplus detached in the first 4 h post-treatment, and 84% detached within the first 24 h. All of the treatments, except the lowest concentration, provided protection of cattle against re-infestation by B. microplus larvae for 14 days post-treatment. Possibly as a result of the formation of a compact layer of lime and amitraz on the bottom after the vat was undisturbed for six weeks, intense agitation was required to re-suspend the active ingredient.

Animals↗

Therapeutic and persistent efficacy of fipronil against Boophilus microplus (Acari: Ixodidae) on cattle.

Cattle infested with all parasitic life-stages of Boophilus microplus (Canestrini) were treated once with a pour-on formulation of fipronil at 0.25, 0.5, or 1.0% active ingredient (AI) to determine the therapeutic efficacy. Additionally, cattle were infested at weekly intervals for 13 consecutive weeks after treatment to determine the persistent effectiveness of the three concentrations. Results of therapeutic efficacy of fipronil showed that with each increase in concentration there was a corresponding and often significant (P < 0.05) decrease in the numbers, fecundity and fertility of engorged females. The overall control of the Index of Reproduction (IR) achieved at 0.25 and 0.5% AI was 86.2 and 94.3%, respectively, while the control of the IR at the 1.0% AI concentration was 99.7%. The persistent efficacy of fipronil, like the therapeutic efficacy, was also dose related. The 0.25% AI treatment failed to provide complete protection (100%) against larval reinfestation, although control of the IR remained high (> 95%) for the first 6 weeks after treatment. In contrast, at 0.5% AI the control of the IR remained at 100% for the first 4 weeks after treatment, and was > 90% for an additional 2 weeks (6 weeks posttreatment). The 1.0% AI concentration afforded the longest residual activity, providing virtually 100% protection against larval reinfestation for 8 weeks after treatment was applied. Based on the results, neither the 0.25 or 0.5% AI concentration completely eliminated the immature stages of the tick (nymphs and larvae) on the host at the time of treatment (therapeutic efficacy), and the residual activity (persistent efficacy) did not prevent reinfestation by larvae for long periods after treatment. While the 1.0% AI concentration provided both therapeutic and residual efficacy that would be suitable for use in the eradication program, caution is still warranted because a few engorged females detached for up to 25 days after treatment, although none reproduced after the 6th day.

Administration, Topical↗