DNA Fragment Encoding Human IL-1β 163–171 Peptide Enhances the Immune Responses Elicited in Mice by DNA Vaccine against Foot-and-Mouth Disease

DNA vaccine has been tested for protection against foot-and-mouth disease. However, the relatively low efficacy of DNA vaccine in inducing immune responses in large animals has restricted its practical use. Interleukin-1 plays an essential role in amplifying both the cellular and humoral immune responses to foreign antigens, and may therefore represent a good candidate as an adjuvant of DNA vaccines. Since the inflammatory activity of IL-I may restrict its application in DNA vaccine treatment, we explored the possibilities of augmenting immune responses without unwanted inflammatory effects using the IL-1beta fragment (amino acids (aa) 163-171), which is essential for IL-1 receptor-1 binding. The DNA fragment encoding the human IL-1beta fragment (aa 163-171) was fused to foot-and-mouth disease virus (FMDV) DNA vaccine, and injected into mice to analyse its immune response. Compared with control mice receiving FMDV DNA vaccine alone, significant increases in the FMDV-specific antibody response and also in T cell proliferation were observed in mice receiving IL-1beta (163-171)-FMDV. These results suggested that DNA fragment encoding IL-1beta 163-171 peptide might represent a good candidate for an adjuvant of FMDV DNA vaccine.     

Acute and Subacute Metabolic and Endocrine Effects of Clenbuterol in Female Pigs

The aim of the study was to assess the relationship between acute and subacute metabolic and endocrine effects after intravenous administration of the ₂-adrenergic agonist clenbuterol in a growth-promoting dose to female pigs. Acute metabolic and endocrine effects were assessed by measuring the blood glucose, serum insulin and nonesterified fatty acid (NEFA) concentrations during 300 min after a single administration of clenbuterol. Significantly higher serum insulin and NEFA concentrations (19.90±2.50 U/ml, p<0.01, and 0.69±0.04 mmol/L, p<0.001, respectively) were measured 30 min after the preprandial administration of clenbuterol in female pigs. Over the same period, the levels of blood glucose (4.42±0.30 mmol/L) showed no difference from those of control pigs. The postprandial serum NEFA concentration decreased moderately during 210 min after feeding. Postprandial blood glucose and insulin concentrations increased and reached maximal levels 120 min after clenbuterol administration (10.91±0.60 mmol/L and 85.22±7.24 U/ml, respectively), and returned to basal levels at 300 min (4.20±0.21 mmol/L and 7.75±1.60 U/ml, respectively) after the administration of clenbuterol. Subacute metabolic and endocrine effects were assessed by measuring the blood glucose, serum insulin and NEFA concentrations for 21 days after the repeated doses of clenbuterol. In addition, the influence of clenbuterol administration on the endocrine regulation of the onset of the next expected oestrus in female pigs was assessed by measuring their serum 17-oestradiol and progesterone concentrations. Blood glucose, serum insulin and NEFA concentrations after the last administration of clenbuterol did not differ significantly from those in control animals. The onset of the next expected oestrus occurred regularly without any significant difference in serum 17-oestradiol or progesterone concentrations between the treated (9.83±2.60 pg/ml and 0.15±0.03 ng/ml) and control pigs (8.52±2.70 pg/ml and 0.25±0.06 ng/ml). The study results suggest the duration of intravenous administration of clenbuterol in a growth-promoting dose necessary to influence the metabolic and endocrine activities in female pigs.     

Reduced Ovulatory and Oestrous Activity in Zebu Heifers Following Trypanosoma vivax Infection

This paper describes a study on the oestrous and ovarian activity and responses to prostaglandin F2 (PGF2) administration and artificial insemination in zebu heifers.Four cycling heifers were artificially infected with 5×106 Trypanosoma vivax organisms. Two heifers served as controls. Two injections of PGF2 were given 11 days apart, commencing at the peak of parasitaemia in the infected animals, followed by artificial insemination 72 and 96 h after the second administration of PGF2. Sera were analysed for progesterone by radioimmunoassay, while ovarian activity and oestrus were determined by rectal palpation and visual observation, respectively. All the infected heifers developed the clinical disease. All control and infected heifers had progesterone profiles consistent with luteolysis and the occurrence of oestrus following the second administration of PGF2. Progesterone levels did not return to normal luteal values in infected animals, however, whilst they did so in control animals. No control or infected heifers became pregnant.The findings suggest that PGF2 will induce a non- fertile oestrus in zebu heifers acutely infected with T. vivax. Re-ovulation is also inhibited within 22 days in a majority of infected animals.     

Gentamicin Nephrotoxicity – A Comparison of In Vitro Findings with In VivoExperiments in Equines

The aminoglycoside gentamicin is often used in equine practice. Despite its clinical use, concerns remain regarding the potential toxic side-effects, such as nephrotoxicity, in equine patients, particularly after repeated dosing. The aim of the study was to investigate first in vitro the mechanisms contributing to the renal toxicity of gentamicin and to identify sensitive biomarkers indicating proximal tubule damage. To this end, the kidney-derived cell lines LLC-PKI and MDCK were treated with gentamicin at different concentrations. Toxicity was assessed by measuring the release of gamma-glutamyl transferase (GGT), and the production of reactive oxygen species (ROS). Cell viability was measured using Alamar blue (AB) and Neutral red (NR) cytotoxicity assays. Gentamicin exerted a dose-dependent toxicity. Primarily, loss of brush border membrane integrity, indicated by GGT leakage, and an increased ROS production were observed. As GGT was found to be a sensitive marker for gentamicin-induced renal cell injury, in the subsequent in vivo experiments, in which ponies were given gentamicin (3.0 mg/kg bw three times daily and 4.5 mg/kg bw twice daily) for five consecutive days, plasma levels and the urinary excretion of GGT and creatinine were measured and the GGT:creatinine ratio was calculated. Elevated GGT levels in urine following gentamicin therapy were observed, but this enzyme leakage was transient and returned to baseline values after cessation of therapy. It could thus be concluded that even a conservative dose regimen of gentamicin did not result in significant renal toxicity in healthy ponies.     

Residues of clenbuterol in tissues of the rainbow trout (Oncorhynchus mykiss)

There have been many studies on the efficacy of ₂-adrenergic drugs as feed additives but no data are available at present on the use of clenbuterol in fish production. To evaluate the residues of clenbuterol in tissues of fish, 50 trout (Oncorhynchus mykiss) were fed for 21 days on a fish feed containing 5 ppm of the drug. The livers, muscles and skins of sample groups of fish were analysed by HPLC with visible spectrophotometric detection on days 15 and 21 of treatment and at intervals during a 30-day withdrawal time. Clenbuterol reached its highest levels in the liver (mean 440 ppb; SD=±159;n=5) on day 15 of treatment, with a slow depletion curve; 24±3 ppb was still present at the end of the withdrawal period. At this time, residues were still present in the edible tissues, i.e. muscle (5±1 ppb) and skin (7±3 ppb). Side-effects were noted during the first week of treatment.Abbreviations ppb parts per billion - BHT butylated hydroxy toluene - CV coefficient of variation - DAD diode array detector - HPLC high-performance liquid chromatography     

Muscarinic receptor subtypes, β-adrenoceptors and cAMP in the tracheal smooth muscle of conventional and double-muscled calves

The total muscarinic (M₁ + M₂ + M₃) and -adrenergic receptors in the tracheal smooth muscle of conventional and double-muscled calves were identified and characterized with the non-specific antagonists [³H]quinuclidinyl benzilate ([³H]QNB) and [³H]dihydroalprenolol ([³H]DHA) respectively.Although the quantity of -adrenoceptors in double-muscled calves was 25% lower (p<0.05) than in conventional calves (B _max=327±89 fmol/mg protein), adenylate cyclase assays indicated that the basal adenylate cyclase activity and the (–)-isopropylnoradrenaline (ISO)- and sodium fluoride (NaF)-stimulated values were not significantly different between these calves. However, the density of muscarinic receptors in double-muscled calves was 40% higher (p<0.01) than in conventional calves (B _max=2955±625 fmol/mg protein). Subtypes of muscarinic receptors were studied with [³H]telenzepine (M₁-receptors), [³H]AF-DX 384 (M₂-receptors) and [³H]4DAMP (M₁ and M₃-receptors). It was found that in both double-muscled and conventional calves about 40% of the receptors were of the M₃-subtype, the remaining 60% being M₂-receptors.From these results, it is suggested that inflammation of the respiratory tract in double-muscled calves may be complicated by an imbalance between the cholinergic bronchoconstrictor and the -adrenergic bronchodilator components of the autonomic nervous system.     

A preliminary study on the effects of atropine sulphate on bradycardia and heart blocks during romifidine sedation in the horse

Romifidine (STH 2130-Cl or Sedivet) is an ₂-agonistic imino-imidazol sedative for intravenous use in horses recently developed by Boehringer Ingelheim, Vetmedica GmbH. An exploratory study was done in nine warm-blood horses, randomly divided into three groups, which received different dosages of romifidine (0.04, 0.08 and 0.12 mg/kg of body weight (BWT) intravenously (i.v.)) with at least one week's interval between tests.Romifidine induced a marked bradycardia accompanied by second degree atrioventricular (AV) block and some sinus blocks at all tested dosages. A placebo (NaCl 0.9% i.v.) given 5 min before and after romifidine did not affect the cardiac disturbances induced by romifidine.A low dose of atropine sulphate (0.005 mg/kg of BWT i.v.) given 5 min before romidifine counteracted the bradycardia and caused a normal to increased heart rhythm at all romifidine dosages. A higher dose of atropine sulphate (0.01 mg/kg of BWT i.v.) administered 5 min before sedation induced a tachycardia (average 70 beats/min) at all romifidine dosages and completely prevented the bradycardia and the heart blocks. The positive chronotrope effects of atropine sulphate were attenuated by increasing doses of romifidine.The effects of atropine sulphate (low or high doses) given 5 min after romifidine only appeared after 5 min. Both dosages counteracted the bradycardia and suppressed the heart blocks.No atropine-dependent side effects were observed in non-fasted horses. The degree of the romifidine induced sedation was not affected by the use of atropine sulphate given before or after romifidine.     

Preliminary findings of altered follicular activity in Holstein cows with coagulation factor XI deficiency

Factor XI (F XI) deficiency is an autosomal recessive coagulopathy found in Holstein cattle. Affected animals have a 50% greater prevalence of repeat breeding. Therefore, several parameters describing ovarian function were studied. Daily blood sampling revealed that progesterone concentrations were slower to decline from a peak at day 16 (p<0.01) to values less than 3 nmol/L in F XI-deficient cows (5.14±0.69 days (mean ± SD) versus 4.05±0.63 days in control animals), resulting in an oestrous cycle length of 24.7±2.1 days compared to 22.9±3.0 days, respectively. This was not due to an alteration in the availability of prostaglandin F₂ (PGF₂) or oxytocin (OT) involved in luteolysis. No significant differences (p>0.05) were seen between normal (n=7) and F XI-deficient (n=7) cows in the peak values or the area under the curve for the pulse in 13,14-dihydro-15-keto PGF₂ in response to OT challenge or in the parameters describing the pulse of ovarian OT secretion after PGF₂ injection (n=7 for each) between days 12 and 14. Ovulatory follicular development was assessed by ultrasound monitoring and plasma 17-oestradiol values at 8-h intervals after a luteolytic injection of cloprostenol (n=6 for each). Follicular diameter was smaller (p<0.05) and accompanied by lower peak oestradiol values near the time of ovulation in F XI-deficient cows. The results suggest that the oestrous cycle in F XI-deficient cows is characterized by a slower process of luteolysis that may be associated with smaller follicular development.Abbreviations F XI factor XI - OT oxytocin - PGF₂ prostaglandin F₂ - PGFM 13,14-dihydro-15-keto-prostaglandin F₂ - i.m. intramuscularly     

Detection of β2-agonists in milk replacer

₂-Agonist drugs may be illegally used as growth promoters for feedlot calves, when mixed into milk replacer immediately before feeding. To check for the presence of clenbuterol, salbutamol and terbutaline in such food, an analytical system was established using a screening method based on two commercial qualitative competitive ELISA tests, with antibodies raised against the arylamino group and thet-butyl group. The extraction procedure was based on precipitation of the milk samples with acetonitrile followed by filtration. The absence of any significant interference by other substances in the filtrate allowed detection of ₂-agonist drugs in spiked samples at the lowest concentration having a repartitioning effect (50 ppb for clenbuterol, mabuterol and terbutaline, 500 ppb for salbutamol). In view of a false positive response with tetracycline in milk samples and a cross-reaction between clenbuterol and mabuterol, an HPLC-MS technique was developed which, after extraction and purification of the samples with SPE C18 Polar Plus, was able to confirm the presence of these drugs. The good recovery after extraction (ranging from 84% to 90.2%) and the low detection limit with this method (250 ng/ml for clenbuterol, mabuterol and terbutaline, and 2.5 µg/ml for salbutamol) allowed easy confirmation and simultaneous detection of the four ₂-agonists at the lowest concentrations at which they are used in adulterated milk for calves.Abbreviations B optical density of the sample - B maximal optical density in total absence of competition - %B/B ₀ percentage of inhibition - ELISA enzyme-linked immunosorbent assay - EIA enzyme immunoassay - HPLC-MS high-performance liquid chromatography-mass spectrometry - m/z mass to charge ratio - ppb parts per billion - ppm parts per million - SPE solid-phase extraction     

Observations on the effects of long-term withdrawal on carcass composition and residue concentrations in clenbuterol-medicated cattle

The detection of the illegal use of clenbuterol (CBL) as a growth promoter has relied on detecting residual concentrations of the drug in body fluids or tissues. Analysis of retinal extracts has recently been shown to considerably extend the detection period following withdrawal. The withdrawal periods required to eliminate residues from the liver and retina were investigated by medicating 20 cattle with CBL for 30 days; 6 control animals remained unmedicated. Residual concentrations were monitored throughout this period and for the subsequent 140 days. Concurrent changes in muscle areas and backfat thicknesses were recorded by ultrasound.CBL was detectable in liver up to the 56th day of withdrawal (0.35 ng/g, SD=0.5), but retinal concentrations remained well above detectable concentrations throughout the withdrawal period (22.5 ng/g, SD=6.5). There were small gains (3–4%) in the muscle areas of treated cattle during medication as compared to controls (p>0.05). These comparative gains remained during withdrawal. Backfat thicknesses in treated animals were 40% lower than in controls at the end of medication (p<0.01). However, by 70 days after withdrawal this difference had disappeared (p>0.05) owing to accelerated fat deposition in the treated group. The retina has been shown to be a highly effective target matrix for detecting CBL administration after long withdrawal periods.Abbreviations CBL clenbuterol - CIM cimaterol - EC European Community - EIA enzyme immunoassay - GC-MS gas chromatography-mass spectrometry - HRPo horseradish peroxidase - L. dorsi, Longissimus dorsi - MRL maximum residue level - SD standard deviation - SED standard error of the difference     

Identification of Functional α-Adrenoceptor Subtypes in the Bovine Female Genital Tract During Different Phases of the Oestrous Cycle

The concentration and functionality of the -adrenoceptor (-AR) subtypes in the genital tract of cyclic heifers were investigated. In each tissue sample, a single class of ₁-ARs was observed, whereas two distinct classes of ₂-ARs were discriminated: low-affinity (LA) and high-affinity (HA) ₂-ARs. Statistical analysis showed the presence of significantly (p < 0.05) higher concentrations of all -AR subtypes in the follicle than in the corpus luteum. No significant differences were found in the ovary or myometrium between the luteal and follicular phases. In the ovary, the density of ₁-ARs was significantly (p < 0.05) higher than that of ₂-ARs. By contrast, there were significantly (p < 0.05) more ₂-ARs than ₁-ARs in the myometrium. As far as ₂-ARs are concerned, LA ₂-ARs were significantly (p < 0.05) higher than HA ₂-ARs in all tested tissues. Competition studies suggested that the rank order of potency of antagonists for ₁-ARs was prazosin > phentolamine > yohimbine, whereas for ₂-ARs the order of potency was yohimbine phentolamine > prazosin. Functional assays performed on myometrium showed that noradrenaline, phenylephrine and clonidine elicited concentration-dependent contractions only in dioestrus and pro-oestrus preparations and that clonidine was more effective than phenylephrine as a contractile agent. It appeared that there were no significant modifications in -AR affinity or concentration during the different stages of bovine oestrous cycle, whereas the uterine spontaneous activity and the responsiveness to -adrenergic stimulation was strongly influenced by hormonal levels. The modifications of uterine contractility observed during the oestrous cycle may be related to modifications induced in the transductional mechanisms of -ARs.     

Effect of Gossypol on Hepatic and Serum γ-glutamyltransferase Activity in Rats

A single intraperitoneal dose (25 mg/kg) of gossypol given to male Sprague-Dawley rats caused marked changes in the activity of the hepatic and serum -glutamyltransferase (GGT) and microsomal monooxygenases. The GGT activity in liver homogenate, S-9 supernatant fraction and microsomes was significantly depressed; however, the level of serum GGT was elevated. While the hepatic glutathione concentration was not greatly changed, the aminopyrine N-demethylase activity and microsomal cytochrome P450 content of the liver were significantly decreased in the treated rats. At necropsy, the livers of the treated rats appeared generally pale with distinct pinpoint foci. Histopathological examination of the liver showed degenerative changes and coagulative necrosis. The results indicate that gossypol is a strong hepatotoxic agent which can produce severe hepatic damage.     

Training-induced modifications in some biochemical defences against free radicals in equine erythrocytes

Oxidative stress develops when the generation of free radicals exceeds the antioxidant capacity of cells or extracellular fluids. It can also occur as a result of physical exercise, and the pathogenesis of exercise-induced myopathies and haemolysis in horses may be related to changes in lipid peroxidation caused by free radicals. Cells have developed biochemical protection against oxidative stress and, as tissues seem to increase their antioxidant defences under chronic activation, training may be one of the ways of increasing antioxidant defences. Accordingly, we tested some enzymatic antioxidant activities as well as nonenzymatic antioxidants in horses undergoing special training. The results indicated a decrease in both chemical and biochemical defences against free radicals during training. It was deduced that the horses' diet may have been unable to provide the increased need for antioxidant defences resulting from training.Abbreviations DPH 1,6-diphenyl-1,3,5-hexatriene - EU enzymic unit - GSH-Px glutathione peroxidase - P degree of fluorescence polarization - Phospholipid P phosphorous content in phospholipids - RBC red blood cell - t-ButOOH t-butylhydroperoxide     

Bovine Immunodeficiency Virus Expression in Vitro is Reduced in the Presence of β-Chemokines,MIP-1α, MIP-1β and RANTES

The inhibition of HIV expression in vitro by a cocktail of the -chemokines MIP-1, MIP-1 and RANTES provided the initial evidence that HIV utilizes chemokine receptors as co-receptors for infection of cells. Bovine immunodeficiency virus (BIV), a lentivirus, infects a wide variety of leukocyte populations, but the cellular receptor(s) utilized by this virus for infection of cells is not known. The purpose of this study was to determine whether MIP-1, MIP-1 and RANTES affect BIV expression in vitro, as a prelude to identifying the cellular receptors utilized by this virus. Fetal bovine lung (FBL) cells were pretreated with serial dilutions of a cocktail of the chemokines, and then the cells were infected with BIV. Virus expression in these cells was determined by counting the syncytia that had developed in the cultures by five days after infection. A significant decrease in syncytium formation, corresponding to increasing concentrations of the chemokines, was the result. Reacting the chemokines with chemokine-specific neutralizing antibodies prior to treatment of the cells neutralized the effect of the chemokines on virus replication in a dose-dependent manner, restoring viral expression to a level similar to that of untreated cells. The presence of a CCR5 homologue on the surface of FBL cells was confirmed using an anti-CCR5 monoclonal antibody and FACS analysis. Collectively, these data provide preliminary evidence that BIV may utilize the CCR5 receptor for infection of cells in vitro, but additional studies are necessary to confirm this.     

Fertility of N'dama and Bunaji Cattle to Artificial Insemination Following Oestrus Synchronization with PRID and PGF2α in the Hot Humid Zone of Nigeria

A study was undertaken to determine the effectiveness of a progesterone-releasing intravaginal device (PRID) and prostaglandin F2 alpha (PGF2alpha) in synchronizing oestrus in N'dama and Bunaji cows and heifers and the fertility following artificial insemination at the synchronized oestrus. A total of 116 cows and heifers (58 N'dama and 58 Bunaji) were used in two separate trials. In the first trial, oestrus was synchronized using a PRID, which was inserted for 12 days; in the second trial, oestrus was synchronized by giving two injections of PGF2alpha 13 days apart. Only animals that did not respond to the first injection were given the second injection. At the end of each treatment period, the animals were observed for oestrus for 7 days and inseminated approximately 12 h following detection of oestrus. Standing to be mounted was the single criterion used to judge an animal to have been in oestrus. PGF2alpha and PRID were both effective in synchronizing oestrus in N'dama and Bunaji cows and heifers. The respective oestrus response rates, pregnancy rate and conception rates for PRID and PGF2alpha were 85.7%, 53.6% and 62.5% for PRID, and 91.7%, 68.3% and 74.6% for PGF2alpha. N'dama cattle showed significantly (p<0.05) better oestrus response rate, pregnancy rate and conception rate than Bunaji cattle following both PRID and PGF2alpha treatments. The pregnancy rate and conception rate following PGF2alpha treatment were better (p < 0.05) than for PRID, although the oestrus response rate did not differ. It is concluded that both PRID and PGF2alpha are effective in synchronizing oestrus in N'dama and Bunaji cattle in the hot humid zone of Nigeria and the fertility to artificial insemination at the synchronized oestrus was normal and acceptable. Thus, PRID and PGF2alpha can effectively be used in intensive breeding programmes for the rapid multiplication and distribution of both cattle breeds, especially the N'dama, which is a unique and beneficial animal genetic resource for the tsetse infested hot humid zone of Nigeria.     

A Comparison of the Concentrations of C-reactive Protein and α1-Acid Glycoprotein in the Serum of Young and Adult Dogs with Acute Inflammation

The concentrations of C-reactive protein (CRP) and ₁-acid glycoprotein (AAG) were evaluated in 1-, 3- and 18-month-old dogs (four of each age) that had been inoculated with turpentine oil. The CRP and AAG in 3-month-old and younger dogs subjected to surgery or inoculated with either Staphylococcus aureus or a viral vaccine were also evaluated. The average CRP concentration in the sera peaked 2 days after inoculation of turpentine oil. The peak CRP concentrations in 3- and 18-month-old dogs were significantly (p<0.05) greater than those in 1-month-old dogs. The average AAG concentration in the sera peaked 4 days after inoculation of turpentine oil. No significant difference was found in AAG concentrations between any of the age groups. When experimentally inoculated with S. aureus or subjected to oophorohysterectomy, the CRP and AAG concentrations increased in 3-month-old dogs, but they increased little in 1-month-old dogs. The CRP and AAG in dogs inoculated with the viral vaccine did not increase. In dogs with fractures or subjected to percutaneous gastrostomy, the CRP and AAG concentrations correlated with the condition of dogs.     

Applications of DNA amplification techniques in veterinary diagnostics

An overview of the principles of the polymerase chain reaction, ligase chain reaction, self-sustained sequence replication and Q replicase is given. The application of these methods for the diagnosis of veterinary infectious and hereditary diseases as well as for other diagnostic purposes is discussed and comprehensive tables of reported assays are provided. Specific areas where these DNA-based amplification methods provide substantial advantages over traditional approaches are also highlighted. With regard to PCR-based assays for the detection of viral pathogens, this article is an update of a previous review by Belák and Ballagi-Pordány (1993).These two authors contributed equally to this review and are listed in alphabetical order.     

Hydrogen Peroxide Production by Mycoplasma bovis and Mycoplasma agalactiae and Effect of In Vitro Passage on a Mycoplasma bovis Strain Producing High Levels of H2O2

Hydrogen peroxide (H2O2) production and oxygen uptake during the oxidation of NADH and L-alpha-glycerophosphate (GP) by lysed cells was determined for the type and field strains of Mycoplasma bovis and M. agalactiae. NADH oxidation by all the strains showed variable production of H2O2 ranging from 0 to 1.21 mol/mol O2 taken up. All strains were unable to oxidize GP, showing absence of GP oxidase activity. Some strains were identified that produced relatively high levels of H2O2 (> 1.0 mol/ mol O2 taken up). In vitro passage of M. bovis strain 119B96 showed reduced H2O2 production: 0.52, 0.16, and 0.07 mol/mol O2 taken up after the 50th, 100th and 200th passages, respectively. SDS-PAGE analysis showed the loss of a protein band of 32 kDa after 50 passages. These preliminary studies show that not only does H2O2 production by potentially pathogenic Mycoplasma spp. vary in the field but also that similar alterations can be induced by passage in culture. In the latter case, at least in one M. bovis strain, this alteration has been shown by SDS-PAGE to be associated with a loss of specific protein production. Further study of these phenomena is essential background for the production of more efficient vaccines for mycoplasmas.