Reversal of profound rocuronium block monitored in three muscle groups with sugammadex in ponies.

BACKGROUND: This clinical study evaluated the speed of reversal of profound rocuronium block in ponies using sugammadex and investigated the differences in onset and recovery from block in three different muscle groups. METHODS: Anaesthesia was induced and maintained with isoflurane in oxygen 100% in eight ponies. Neuromuscular monitoring was performed at each site using acceleromyography: in the extensor muscles of the pelvic limb (peroneal nerve) and thoracic limb (radial nerve), and in the orbicularis oris muscle (OOM; facial nerve). Rocuronium 0.6 mg kg(-1) i.v. was administered, followed 5 min later by sugammadex 4 mg kg(-1) i.v. Onset time (onsetROC), maximum block, and time to recovery of the train-of-four ratio to 0.9 (TOFR=0.9) were recorded. The differences between monitored sites were compared using one-way anova followed by a post hoc Dunn's test. RESULTS: Onset of ROC was significantly delayed in OOM compared with both limbs [pelvic limb, thoracic limb, and OOM: 43.1 (sd 16.9), 50.6 (15.9), and 204.4 (35.8) s, respectively; P<0.001]. Complete block was achieved in the pelvic and thoracic limbs, but in none of the eight ponies in the OOM [mean T1=15.3 (9.4)%; range: 7-36%]. No differences were observed between muscle sites in recovery to TOFR=0.9 [pelvic limb, thoracic limb, and OOM: 2.3 (0.9), 3.4 (1.7), and 2.8 (2.1) min, respectively]. No adverse effects of sugammadex were detected throughout the study period. CONCLUSIONS: Sugammadex can be used to reverse profound rocuronium-induced block in ponies during isoflurane anaesthesia. Thoracic limb muscles represent a suitable alternative for monitoring neuromuscular block compared with pelvic limb muscles.

A cadaver study comparing two approaches to perform a maxillary nerve block in the horse.

REASONS FOR PERFORMING STUDY: Anaesthesia of the maxillary nerve of the horse has been described using several approaches, but sparse data exist to evaluate the accuracy of these methods. OBJECTIVES: This study compared 2 previously described approaches to the maxillary nerve to assess their relative accuracies. METHODS: Thirty severed heads from horse cadavers were arranged to approximate the position of a live horse. Methylene blue (0.25 or 0.1 ml) was injected using a 19 gauge 90 mm spinal needle by one of 2 approaches, the method used being randomly allocated in each instance. Method ANG: angulated needle insertion on the ventral border of the zygomatic process of the temporal bone and directed rostromedially. Method PER: needle inserted perpendicular to the skin surface, ventral to the zygomatic process of the malar bone, level with the temporal canthus of the eye. Accuracy of dye deposition was assessed following dissection. Placement was categorised as 'full hit' (complete nerve coverage or dye deposition centred on nerve), 'partial hit' (partial nerve discolouration but dye not centred on nerve) or 'miss' (no nerve discolouration). Deposition of dye relative to the nerve and whether injection was performed on the left or right side of the head was recorded. A Chi-squared test was performed to examine the relationship between the 2 methods. RESULTS: Method ANG was performed 31 times, Method PER 28 times. Full hits were 10/31 (32%) vs. 9/28 (32%), partial hits 15/31 (49%) vs. 14/28 (50%) and misses 6/31 (19%) vs. 5/28 (18%) (Methods ANG vs. PER, respectively). RESULTS were not statistically significantly different between the methods. Dye was deposited in the deep facial vein once by each method. Bone was contacted consistently with Method PER and 8/31 times with Method ANG. CONCLUSION AND CLINICAL RELEVANCE: Both methods appeared equivalent in terms of accuracy. Aspiration should always precede injection.

Restoration of arterial oxygen tension in horses recovering from general anaesthesia.

BACKGROUND: Arterial hypoxaemia is common in anaesthetised horses, but little information exists regarding restoration of arterial oxygen tension (PaO2 ) during recovery from anaesthesia, or if intra-operative management factors exert any longer-term effect. OBJECTIVES: To evaluate PaO2 in horses recovering from general anaesthesia up to 1 h after resuming standing. STUDY DESIGN: Prospective observational clinical cohort study. METHODS: Systemically healthy adult horses undergoing inhalational general anaesthesia for elective surgical procedures were studied. Arterial blood samples were obtained anaerobically prior to pre-anaesthetic medication, at end of anaesthesia, immediately following positioning in the recovery box, then at 10-min intervals until standing. Additionally, samples were taken when horses achieved sternal recumbency, at standing (STAND) and 1 h after standing (STAND+1). Data were analysed using ANOVA and mixed-effects linear regression, with significance set at P<0.05. RESULTS: Data from one hundred and two horses were analysed. Forty horses received controlled mandatory ventilation (CMV) throughout anaesthesia, 47 breathed spontaneously (SV) and 15 initially breathed spontaneously before CMV was imposed (S-CMV). Overall, PaO2 , P(A-a)O2 and PaCO2 remained significantly lower than baseline at STAND+1 (P<0.01). CMV resulted in higher PaO2 at the end of anaesthesia (P = 0.03) and during early recovery (P<0.01) than SV. Only in group S-CMV did PaO2 , P(A-a)O2 and PaCO2 return to baseline values at STAND+1. Highest PaO2 values associated with CMV were also associated with early recovery apnoea. MAIN LIMITATIONS: Non-standardised anaesthetic management, temporal and quantitative variation in oxygen delivery during early recovery and lack of control group where oxygen was electively withheld during recovery. CONCLUSIONS: Controlled mandatory ventilation results in better pulmonary function in horses as assessed by PaO2 , P(A-a)O2 and PaCO2 , an effect enhanced by an initial period of SV and still evident 1 h after standing. High PaO2 values may contribute to early recovery apnoea but this does not adversely affect outcome. The Summary is available in Portuguese - see Supporting Information.

Differences in plasma and peritoneal fluid proteomes identifies potential biomarkers associated with survival following strangulating small intestinal disease.

BACKGROUND: Strangulating small intestinal disease (SSID) carries a poor prognosis for survival in comparison to other types of colic, particularly if resection is required. Identification of markers which aid early diagnosis may prevent the need for resection, assist with more accurate prognostication and/or support the decision on whether surgical intervention is likely to be successful, would be of significant welfare benefit. OBJECTIVES: To apply an unbiased methodology to investigate the plasma and peritoneal fluid proteomes in horses diagnosed with SSID requiring resection, to identify novel biomarkers which may be of diagnostic or prognostic value. STUDY DESIGN: Prospective clinical study. METHODS: Plasma and peritoneal fluid from horses presented with acute abdominal signs consistent with SSID was collected at initial clinical examination. Samples from eight horses diagnosed with SSID at surgery in which resection of affected bowel was performed and four control horses subjected to euthanasia for orthopaedic conditions were submitted for liquid chromatography tandem mass spectrometry. Protein expression profiles were determined using label-free quantification. Data were analysed using analysis of variance to identify differentially expressed proteins between control and all SSID horses and SSID horses which survived to hospital discharge and those which did not. Significance was assumed at P≤0.05. RESULTS: A greater number of proteins were identified in peritoneal fluid than plasma of both SSID cases and controls, with 123 peritoneal fluid and 13 plasma proteins significantly differentially expressed (DE) between cases and controls (P<0.05, ≥2 fold change). Twelve peritoneal fluid proteins (P<0.036) and four plasma proteins (P<0.05) were significantly DE between SSID horses which survived and those which did not. MAIN LIMITATIONS: A low number of samples were analysed, there was variation in duration and severity of SSID and only short-term outcome was considered. CONCLUSIONS: Changes in peritoneal fluid proteome may provide a sensitive indicator of small intestinal strangulation and provide biomarkers relevant to prognosis.

Clinical Research Abstracts of the British Equine Veterinary Association Congress 2015.

REASONS FOR PERFORMING STUDY: Colic remains a life-threatening condition in the horse. Ischaemia and reperfusion following correction of small intestinal strangulation may produce oxidative stress. The ability to withstand oxidative stress depends on antioxidant levels and may be linked to horse survival. OBJECTIVES: To measure peripheral antioxidant levels in horses undergoing exploratory laparotomy with small intestinal strangulation. STUDY DESIGN: Case-control study. METHODS: Blood and plasma were collected from horses undergoing exploratory laparotomy for small intestinal strangulation and stored at -80°C. Controls involved non-colic horses. Total plasma glutathione was measured spectrophotometrically at 412 nm using the 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB, Ellman's reagent) reaction. Samples containing scavenger (to remove reduced glutathione, GSH) were used to measure oxidised glutathione (GSSG). Glutathione reductase (GR) activity (u/l) was measured as the rate of GSH production at 412 nm. Glutathione peroxidise (GPx) activity (u/l) was measured as the change in optical density (340 nm) following the consumption of NADPH after GSSG production. All assays were purchased from BioAssay Systems (Hayward, California). Clinical data including arterial blood gas analysis were collected on admission. RESULTS: Glutathione reductase activity in horses with strangulating small intestinal lesions was significantly reduced compared to control horses (12.2 ± 1.1 u/l vs. 15.9 ± 0.8 u/l, P = 0.03, n = 6) whereas GPx activity did not significantly differ between colic and control horses (155.7 ± 48.7 u/l vs. 167.3 ± 30.1 u/l, P = 0.84, n = 6). Total glutathione, reduced or oxidised glutathione did not differ significantly between control and colic horses. A positive correlation existed between GR activity and Ca(2+) (r = 0.93) and K(+) (r = 0.75) whereas a strong negative correlation was present between GR activity and HCO3 (-) (r = -0.92) and PaCO2 (r = -0.96). CONCLUSIONS: Reduced plasma glutathione reductase activity with small intestinal strangulation indicates oxidative stress and may be related to systemic electrolyte/bicarbonate abnormalities. Ethical animal research: Study approval No. VREC219a. Explicit owner informed consent for inclusion of animals in this study was not stated. SOURCE OF FUNDING: Supported by the School of Veterinary Sciences, University of Liverpool. Competing interests: None declared.

Comparison of the EPOC and i-STAT analysers for canine blood gas and electrolyte analysis.

OBJECTIVES: A prospective study to evaluate agreement and precision of a new point-of-care portable analyser, the EPOC analyser, compared with the i-STAT analyser in canine blood. METHODS: Blood samples (68 venous and 32 arterial) were obtained from 63 client-owned dogs for clinical reasons and surplus blood was used to analyse agreement between the EPOC and i-STAT analysers. Precision of the EPOC analyser was also assessed by repeat analysis of 20 samples. Measured analytes included pH, partial pressures of carbon dioxide and oxygen and concentrations of sodium, potassium, ionised calcium, glucose and haematocrit. Haemoglobin, base excess, bicarbonate and saturation of haemoglobin with oxygen were calculated. RESULTS: EPOC precision was acceptable, but agreement was poor for sodium, haematocrit, haemoglobin and base excess. Overall, method comparison was poor for pH, partial pressure of oxygen, sodium, haematocrit, haemoglobin and base excess. CLINICAL SIGNIFICANCE: The EPOC analyser is useful for dogs, although clinically significant differences between the EPOC and i-STAT analysers exist for some analytes, and as such these analysers should not be used interchangeably.

Factors associated with survival to hospital discharge following endoscopic treatment for synovial sepsis in 214 horses.

REASONS FOR PERFORMING STUDY: To determine risk factors involved in survival to hospital discharge of cases of synovial sepsis. OBJECTIVES: Investigate pre-, intra- and post operative factors involved in short-term survival of horses undergoing endoscopic treatment for synovial sepsis. STUDY DESIGN: Retrospective case series. METHODS: Clinical data were obtained for horses (>6 months old) undergoing endoscopic surgery as part of management for synovial sepsis over a 7-year period in a single hospital population. Descriptive data were generated for pre-, intra- and post operative variables. Multivariable logistic regression analysis was used to develop 3 models related to presurgical, surgical and post surgical stages of management with outcome defined as survival to hospital discharge. RESULTS: Two hundred and fourteen horses were included. In Model 1 (preoperative variables), increased preoperative synovial fluid total protein (TP) was associated with nonsurvival (OR 0.88, 95% CI 0.83-0.94, P<0.001) whereas the presence of a wound on admission was associated with survival (OR 4.75, 95% CI 1.21-18.65, P = 0.02). Model 2 (intraoperative variables) revealed that factors associated with decreased survival were anaesthetic induction outside of normal working hours (OR 0.36, 95% CI 0.15-0.88 P = 0.02) and presence of moderate/severe synovial inflammation at surgery (OR 0.28, 95% CI 0.12-0.67, P = 0.004). Model 3 (post operative variables) showed that increased post operative synovial fluid TP (OR 0.94, 95% CI 0.90-0.98, P = 0.013) and undertaking more than one endoscopic surgery for treatment (OR 0.19, 95% CI 0.05-0.70, P = 0.005) were associated with nonsurvival. Cut-off values for predicting survival were 55-60 g/l for preoperative and 50-55 g/l for post operative TP measurements. CONCLUSIONS: This study has identified factors associated with altered likelihood of survival to hospital discharge following endoscopic surgery for synovial sepsis. Prognosis for survival to hospital discharge can be based on evidence from this study at the key stages of management of horses with synovial sepsis.

Use of continuous positive airway pressure (CPAP) to optimise oxygenation in anaesthetised horses--a clinical study.

REASONS FOR PERFORMING STUDY: Hypoxaemia is a common problem during equine anaesthesia. Continuous positive airway pressure (CPAP) is a ventilation mode routinely employed in man to overcome hypoxaemia but has not been objectively assessed in horses. OBJECTIVES: To test the effects of CPAP on oxygenation and its indices in anaesthetised horses in a clinical setting. METHODS: Twenty-four healthy horses requiring anaesthesia in dorsal recumbency were anaesthetised using a standard protocol. Following orotracheal intubation and connection to an anaesthetic machine capable of applying CPAP, horses were randomly allocated to ventilate at physiological airway pressure measured at the airway opening (Group PAP) or to receive CPAP of 8 cmH2O (Group CPAP). Arterial blood gas analysis was performed as soon as arterial cannulation was achieved and 30, 60 and 90 min after induction. If PaCO2 increased above 9.31 kPa controlled ventilation was initiated. Groups were compared using a general linear model. RESULTS: Horses receiving CPAP had significantly higher PaO2 and calculated oxygen indices than horses receiving PAP. No significant differences in ventilation indices were observed between the 2 groups. Eight horses receiving PAP and 5 receiving CPAP required controlled ventilation. No differences in dobutamine requirements or mean arterial pressures were recorded. CONCLUSIONS: Continuous positive airway pressure of 8 cmH2O improved oxygenation indices in dorsally recumbent horses without significantly influencing ventilation. POTENTIAL RELEVANCE: Continuous positive airway pressure reduces the incidence of hypoxaemia in anaesthetised horses. Further research is warranted to elucidate the effects of CPAP on the cardiovascular system.

Hand-to-hand transmission of herpes simplex virus type 1.

Droplets of tissue culture fluid containing herpes simplex virus type 1 were placed on the palm of the hand. Each 0.01 ml droplet was taken from a stock virus suspension with a titre of 10(7.5) TCID50/0.1 ml. At 0, 15, 30, 60 and 120 min a droplet was firmly touched with the middle finger of the right hand, after which, attempts were made to recover virus from the finger. At 0 min, when the virus-containing droplet was in a liquid state, there was a 100% rate of virus recovery. By 15 min the droplets had dried out, and after touching dried out droplets there was a 40% virus recovery rate, even though experimental procedures demonstrated that infectious virus was present in the dried out droplets at all test times. If the finger was moistened with tap water or saliva, there was a 100% recovery rate of virus after touching dried out droplets, as well as after touching droplets in a liquid state.

Survival of Herpes Simplex Virus Type 1 on Some Common Foods Routinely Touched before Consumption.

Droplets of saliva containing herpes simplex virus type 1 were placed on the skin of tomatoes and the upper surface of lettuce leaves. There was no loss of virus infectivity titer at refrigerator temperature (2°C) at any time examined up to 1 h, the longest period tested. At room temperature (22 to 24°C) there was a 2-log drop in titer between 30 and 60 min, but some infectious virus was still present at 1 h. The virus-containing saliva remained in a liquid state at 2°C. At 22 to 24°C the droplets became dry at approximately 50 min. Implications of the findings are discussed.

Survival of herpes simplex virus type 1 on some frequently touched objects in the home and public buildings.

The survival of herpes simplex virus type 1 in tissue culture fluid on doorknobs and washbasin tap handles over a 2 h period was investigated. The doorknobs were made of plastic and the taps had a chrome-plated surface. The stock virus suspension had a titre of 10(7.5) TCID50/0.1 ml, and a 0.01 ml droplet was placed on the objects. The droplets dried out after 35-40 min at room temperature (23-27 degrees C). The titre of infectious virus recovered by elution from the objects was determined at 0, 15, 30, 60 and 120 min. Infectious virus was recovered for all time periods studied. However, there was a marked drop in titre between 30 and 60 min. No differences were observed between virus placed on plastic and that on chrome-plate. Infectious virus could be isolated from skin after touching virus-contaminated doorknobs and taps with the middle finger of the right hand at all test times over a 2 h period.

Exposure of herpes simplex virus type 1 and its host cells to cigarette smoke in vitro.

A 1.0 ml suspension of herpes simplex virus type 1 in tissue culture growth medium was exposed at 37 degrees C to eight puffs of smoke from one cigarette. Each puff consisted of 25.0 ml of mainstream smoke, and was delivered by a mechanical smoking apparatus. At 0, 15 and 60 min after exposure the virus was assayed for infectivity using HEp-2 cells as the host. Neither filter nor non-filter cigarette smoke affected the infectivity of the virus, or the viability of HEp-2 cells treated with smoke in a manner similar to the virus. The filter cigarette contained 19.0 mg of tar and 1.2 mg of nicotine, and the non-filter cigarette had 23.0 mg of tar and 1.4 mg of nicotine. Smoke from four non-filter cigarettes delivered over a 4 h period caused a 4-log10 drop in virus infectivity titre, and smoke from four filter cigarettes caused more than a 1-log10 drop in titre. Smoke from four non-filter or filter cigarettes was highly cytocidal for HEp-2 cells.

Glucose uptake and lactic acid production of adenovirus type 5-infected HEp-2 cells cultured under exponential growth and stationary phase conditions.

Exponentially growing HEp-2 cells have a higher rate of glucose uptake and lactic acid production than stationary phase cells. Infection of cells with adenovirus type 5 stimulates glycolysis irrespective of the original rate of the host cells. Therefore, infected cells cultured under exponential growth conditions have a higher rate of glycolysis than infected stationary phase cells. The rate of host cell glycolysis does not influence the time required for virus replication, the yield of infectious virus particles produced, or the time of appearance and progression of virus-induced cytopathology.

Differential cell membrane labilizing effect of adenovirus type 5 and 12 observed during productive and abortive infection.

Culture fluid of human epitheloid (HEp-2) cells was examined for extracellular lactate dehydrogenase activity as an indicator of cell damage during a 48 h period in which virus replication and changes in cell morphology occurred. Uninfected and adenovirus type 5-infected cells had the same levels of extracellular enzyme activity both before and after the appearance of morphological changes in cells due to virus infection, whereas adenovirus type 12-infected cells showed increased extracellular enzyme activity. Cells infected with either adenovirus type 5 or type 12 had the same total cellular and extracellular lactate dehydrogenase activity. Hydrocortisone, a membrane stabilizing agent, prevented abnormal leakage of lactate dehydrogenase from adenovirus type 12-infected cells, but had no effect on virus replication or total enzyme activity of infected cells. After inoculation of monkey kidney (Vero) cells the yield of progeny adenovirus type 5 virions was greatly reduced and there was no production of adenovirus type 12 virions. The pattern of extracellular lactate dehydrogenase activity of uninfected and adenovirus type 5- and type 12-infected Vero cells was like that with HEp-2 cells. Therefore, production of adenovirus type 12 virions is not necessary for the virus-cell interaction causing cell membrane labilization.

Viability of six species of normal oropharyngeal bacteria after exposure to cigarette smoke in vitro.

Bacteria in culture medium at 37 degrees C were subjected to eight puffs of smoke from one cigarette, and viable cell counts were done at intervals ranging from 0 min to 3 h after exposure to smoke. The cigarette contained 23.0 mg of tar and 1.4 mg of nicotine. Each 25.0 ml puff of smoke from a mechanical smoking apparatus was passed over a 1.0 ml suspension of bacteria dispersed over a 25 cm2 surface in a sterile flask. Filtered air was in contact with the suspension between puffs, and during the intervals between smoke treatment and viable cell counts being done. There was a marked decrease in numbers of viable Branhamella catarrhalis at 15 min after exposure to smoke, and by 1 h there were no viable cells in suspensions which originally contained 10(7) bacteria. No reduction in the number of viable Branhamella catarrhalis occurred in an untreated suspension of the micro-organism. There was a marked decrease in viable Neisseria perflava at 30 min, and by 3 h there were no viable cells. There was a 4 to 5 log10 reduction of viable Neisseria sicca compared with untreated bacteria at 3 h after exposure to smoke. Streptococcus mitis, Streptococcus salivarius, and Streptococcus sanguis were less susceptible to the harmful action of cigarette smoke, and by 3 h there were 1 to 2 log10 differences in viable cell counts between smoke-treated and untreated bacteria. Non-filter cigarette smoke had a greater detrimental effect on all species of bacteria studied than smoke from filter-tipped cigarettes.

Host cell glucose metabolism during abortive infection by adenovirus type 12.

Progeny virions were not produced following infection of an established line of African green monkey kidney (Vero) cells with human adenovirus type 12. In addition, viral structural proteins were not synthesized as determined by the immunofluorescent procedure to detect those proteins in infected cells. Cytopathology was observed in the virus-infected Vero cells, but it was less pronounced and differed in some other aspects to the cytopathic effect of adenovirus type 12 during productive infection. Glucose uptake and lactic acid production were monitored as indicators of glucose metabolism over a 48 h period during abortive infection. There was a 50 to 60% increase in glucose uptake and lactic acid production by virus-infected Vero cells compared with parallel cultures of uninfected cells.

In vitro exposure of adenovirus types 5 and 7 in oropharyngeal secretions to cigarette smoke.

Adenovirus types 5 and 7 were suspended in clarified oropharyngeal secretions. After 1 ml of suspension was dispersed in a thin layer over a 25-cm2 surface in a flask, the suspension was exposed at 37 degrees C to eight 25-ml puffs of smoke from one cigarette. A mechanical smoking apparatus was used. Nonfilter cigarettes used had 23 mg of tar and 1.4 mg of nicotine, and filter cigarettes used had 19 mg of tar and 1.2 mg of nicotine. Smoke was flushed from the flask with normal filtered air. At 0, 0.25, and 1 h after exposure to smoke, untreated and smoke-treated viruses were titrated with monolayer cultures of human epithelioid (HEp-2) cells. Normal air was in contact with the suspensions between puffs and between smoke treatments and virus titrations. Smoke from filter or nonfilter cigarettes had no effect on the infectivity and replication of adenovirus types 5 and 7. Smoke from four cigarettes administered over a 4-h period caused a 2- to 3-log10 drop in the titers of both viruses. Smoke from four cigarettes was also highly toxic to HEp-2 host cells of the viruses.

Leakage of cellular enzyme during replication of oncogenic and nononcogenic adenoviruses.

Leakage of glutamic-oxalacetic transaminase was monitored as an indicator of cell damage during a 48-hour period in which maximum virus replication occurred. There was increased leakage of the enzyme from HEp-2 cells infected with highly oncogenic adenovirus type 12. Leakage from cells infected with adenovirus types 18 and 31 (highly oncogenic), types 3,7 and 14 (weakly oncogenic), and types 1, 2 and 5 (nononcogenic) was similar to that from uninfected HEp-2 cells, even after the appearance of advanced virus-induced cytopathology.

Replication of adenovirus types 5, 7, and 12 during inhibition of host cell oxidative metabolism.

Replication of human adenovirus type 5 (non-oncogenic), type 7 (weakly oncogenic), and type 12 (highly oncogenic) was studied. Inhibition of cellular oxidative metabolism with sodium cyanide resulted in much lower yields of progeny virions in chimpanzee liver cells, an established cell line derived by biopsy from a normal chimpanzee. Inhibition of oxidative metabolism had no effect on virus replication in HEp-2 cells, an established cell line derived from epidermoid carcinoma tissue from the larynx of a human being. The NaCN, at a concentration of 10(-4) M in cell culture medium, was at a sub-lethal level for host cells during a 48 h period for virus replication under one-step growth conditions.

Studies on the survival and inactivation of herpes simplex virus type 1 on coins.

Survival of herpes simplex virus type 1 (HSV-1) in saliva at room temperature (21-23 degrees C) on 1, 5, 10, and 25 cent coins was studied. There was little or no loss of infectious HSV-1 before 30 min. Between 30 and 60 min there was a 2- to 3-log drop in titre, and with the exception of the 1 cent coin, some infectious virus was still present after 2 h, the longest period studied. The most conspicuous drop in titre occurred with loss of moisture from the saliva. In addition to the drying process, the metals of the coins also contributed to the decline in titre of HSV-1.