Tissue donation and virus safety: more nucleic acid amplification testing is needed.

In tissue and organ transplantation, it is of great importance to avoid the transmission of blood-borne viruses to the recipient. While serologic testing for anti-human immunodeficiency virus (HIV)-1 and -2, anti-hepatitis C virus (HCV), hepatitis B surface antigen (HBsAg), anti-hepatitis B core antigen (HBc), and Treponema pallidum infection is mandatory, there is until now in most countries no explicit demand for nucleic acid amplification testing (NAT) to detect HIV, hepatitis B virus (HBV), and HCV infection. After a review of reports in the literature on viral transmission events, tissue-specific issues, and manufacturing and inactivation procedures, we evaluated the significance of HIV, HCV, and HBV detection using NAT in donors of various types of tissues and compared our results with the experiences of blood banking organizations. There is a significant risk of HIV, HCV, and HBV transmission by musculoskeletal tissues because of their high blood content and the high donor-recipient ratio. If no effective virus inactivation procedure for musculoskeletal tissue is applied, donors should be screened using NAT for HIV, HCV, and HBV. Serologically screened cardiovascular tissue carries a very low risk of HIV, HCV, or HBV transmission. Nevertheless, because effective virus inactivation is impossible (retention of tissue morphology) and the donor-recipient ratio may be as high as 1:10, we concluded that NAT should be performed for HIV, HCV, and HBV as an additional safety measure. Although cornea allografts carry the lowest risk of transmitting HIV, HCV, and HBV owing to corneal physiology, morphology, and the epidemiology of corneal diseases, NAT for HCV should still be performed. If the NAT screening of a donor for HIV, HCV, and HBV is negative, quarantine storage of the donor tissue seems dispensable. In view of numerous synergistic effects with transfusion medicine, it would be advantageous for tissue banks to cooperate with blood bank laboratories in performing virological tests.

Changes in QRS voltage in cardiac tamponade and pericardial effusion: reversibility after pericardiocentesis and after anti-inflammatory drug treatment.

OBJECTIVES: The goal of this study was to define the association between low QRS voltage and cardiac tamponade or pericardial effusion and to assess the reversibility of low QRS voltage after therapeutic procedures. BACKGROUND: It is unclear whether low QRS voltage is a sign of cardiac tamponade or whether it is a sign of pericardial effusion per se. METHODS: In a prospective study design, we recorded consecutive 12-lead electrocardiograms and echocardiograms in 43 patients who were referred to our institution for evaluation and therapy of a significant pericardial effusion. Cardiac tamponade was present in 23 patients (53%). Low QRS voltage (defined as maximum QRS amplitude <0.5 mV in the limb leads) was found in 14 of these 23 subjects (61%). Nine of these 14 patients were treated by pericardiocentesis (group A). Five patients received anti-inflammatory medication (group B). Group C consisted of nine patients with pericarditis and significant pericardial effusion who had no clinical evidence of tamponade. RESULTS: In group A, low QRS voltage remained largely unchanged immediately after successful pericardiocentesis (0.36 +/- 0.17 mV before vs. 0.42 +/- 0.21 mV after, p = NS), but QRS amplitude recovered within a week (0.78 +/- 0.33 mV, p < 0.001). In group B, the maximum QRS amplitude increased from 0.40 +/- 0.20 mV to 0.80 +/- 0.36 mV (p < 0.001) within six days. In group C, all patients had a normal QRS amplitude initially (1.09 +/- 0.55 mV) and during a seven-day follow-up (1.10 +/- 0.56 mV, p = NS). CONCLUSIONS: Low QRS voltage is a feature of cardiac tamponade but not of pericardial effusion per se. Our findings indicate that the presence and severity of cardiac tamponade, in addition to inflammatory mechanisms, may contribute to the development of low QRS voltage in patients with large pericardial effusions.

Assay of antibodies to hepatitis C virus protein C100-3 in blood donors from northern Germany.

The prevalence of anti-C100-3 increases with age from 0.41% to 1.26%. It is more frequent in donors with elevated ALT (4.5%). Most ALT elevations, however, are not related to anti-C100-3. Low EIA signals (< 3 x cutoff) are often non-specific. The cutoff value should be 2.5 times higher. High EIA signals correlate with ALT elevations.

The influence of colostral leukocytes on the course of an experimental Escherichia coli infection and serum antibodies in neonatal calves.

Three hours post natum, 20 calves were orally infected with 10(9) colony forming units of an enteropathogenic E. coli. Immediately after infection and on 2 days subsequent, ten of the calves (COL-) received cell-deprived pooled colostrum and the other ten (COL+) pooled colostrum supplemented with colostral cells. The COL+ calves excreted significantly less bacteria of the infectious strain with their faeces in the first week after infection and reached the lower limit of detectability earlier than the COL- calves. Colostral leukocytes obtained from cows which developed clinical mastitis (n = 2), showed a marked reduction in the number of shed bacteria. The concentration of IgA and IgM specific antibodies against E. coli in the serum of the COL+ calves was significantly higher in the early postnatal period than in the serum of the COL- calves and remained at a slightly higher level throughout the whole investigative period. In addition to humoral substances of the colostrum, colostral leukocytes obviously contribute to the passive immunity and resistance of the newborn calf. The quality and quantity of the leukocytes seem to be of crucial importance to their efficiency.

Uptake of colostral leukocytes in the intestinal tract of newborn calves.

The role of colostral immunoglobulins for the protection of newborn calves has been studied extensively, but little is known about the importance of colostral leukocytes. To study the uptake of colostral leukocytes in the intestine of calves and to determine preferential sites for this uptake, FITC-labelled colostral cells derived from the respective dams were injected into intestinal loops with/without Peyer's patches of three male Holstein Frisian calves about 5h post natum. In adjacent loops, PBS was injected as control. Loops were excised after an exposure of 1.5-2h. FITC-labelled material and cells were detected by the direct immunoperoxidase method in paraplast sections. Twenty-five consecutive sections were evaluated from each localization. Uptake of labelled material and cells was observed in all three calves in the jejunal Peyer's patch and in two calves in the ileal Peyer's patch as well. In the jejunal Peyer's patch, labelled material and cells were present in epithelium, domes and sinuses around lymphoid follicles, whereas in the ileal Peyer's patch, they were found in the sinuses only. These findings confirm that uptake of colostral leukocytes through the intestinal barrier is possible and that the preferential route of uptake is through follicle-associated epithelium of Peyer's patches.

The influence of colostral leukocytes on the immune system of the neonatal calf. III. Effects on phagocytosis.

The influence of colostral leukocytes on the activity of phagocytic cells from the blood of calves, in particular the concentration of neutrophils (PMN) in blood, ingestion of Streptococcus agalactiae, reduction of NBT-dye and activity of lysozyme, was investigated for four weeks using four groups. The calves received either complete colostrum (COL+, n = 16), cell depleted colostrum (COL-, n = 16), cell-supplemented milk-substitute (MS+, n = 7) or pure milk-substitute (MS-, n = 6). Calves of the COL+ group had a significantly lower PMN concentration in their blood on day 2 and a significantly higher activity of lysozyme during their first three weeks of life as compared to the COL- animals. A postnatal increase in number of ingested Streptococcus agalactiae test bacteria per 100 phagocytic cells occurred later in the COL+ calves than in the COL-. No difference between both COL groups in NBT-reduction was observed. The calves of the MS+ group showed higher lysozyme activity and a retarded increase in the ingestion of test bacteria during the first week of life as compared to the MS-. The MS+ group had a transient neutrophilia on the second day of life while the concentration of PMN was not altered in the MS- from the first to the second day.

The influence of colostral leukocytes on the immune system of the neonatal calf. II. Effects on passive and active immunization.

The influence of colostral leukocytes on the concentration of immunoglobulins and antibodies against an enterotoxigenic strain of E. coli in the sera of newborn calves was investigated for four weeks using four experimental groups. The calves received either complete colostrum (COL-, n = 16), cell-supplemented milk substitute (MS+, n = 7) or pure milk substitute (MS-, n = 6) during the first three days of life. The cows were not specifically immunized. The sera of the COL+ calves had significantly higher concentrations of antibodies against E. coli mainly of IgG1 specificity on the second day of life as compared to those of the COL-. The sera of the COL+ calves contained significantly more IgM on days 2 and 5 and slightly more IgA during the first week. Both COL groups had equal concentrations of serum IgG. It appears that colostral leukocytes which are an integral part of the colostrum enhance the passive immunity of the neonatal calf, especially in regard of antibodies and immunoglobulin classes which are essential for intestinal immunity. The concentration of IgM in the sera of the MS+ calves was reduced, that of IgG did not rise to appreciable amounts; the IgA synthesis started one week later as compared to the MS- group. The administration of isolated colostral cells led to an impairment of the natural active immunization.

The influence of colostral leukocytes on the immune system of the neonatal calf. I. Effects on lymphocyte responses.

The influence of colostral leukocytes on lymphocyte counts in the blood of calves and on lymphocyte responses, in particular the Concanavalin A-induced blastogenic response in vitro and the formation of antibodies against sheep erythrocytes, was investigated for four weeks postnatum using four experimental groups. The calves received either complete colostrum (COL+, n = 16), cell-depleted colostrum (COL-, n = 16), colostral cell-supplemented milk substitute (MS+, n = 7) or pure milk substitute (MS-, n = 6) during their first three days of life. In contrast to the calves fed with cell-depleted colostrum (COL-) the calves fed with complete colostrum (COL+) showed no decrease of lymphocyte numbers in the blood on the second day of life, uniform blastogenic responses to two different Concanavalin A concentrations, slightly enhanced antibody formation against sheep erythrocytes and a high spontaneous proliferation of mononuclear cells during the first week of life. In the calves fed with milk-substitute supplemented with colostral cells (MS+) a higher blastogenic response to Concanavalin A and an intensified formation of antibodies against sheep erythrocytes was observed as compared to the MS- calves. A passage of vital colostral lymphocytes through the intestinal wall is postulated. They seem to stimulate and regulate the blastogenic response and enhance the T-helper cell-dependent formation of antibodies against sheep erythrocytes in calves.

[Review article: colostral leukocytes and their significance for the immune system of newborns]. / Ubersichtsreferat: Die Kolostralleukozyten und ihre Bedeutung für das Immunsystem des Neugeborenen.

The colostrum contains a comparably high concentration of leukocytes as the peripheral blood. The majority of them are vital leukocytes, namely neutrophils, macrophages and lymphocytes. There is some evidence in mouse and man that lymphocytes from the gut-associated lymphoid tissue home selectively to the peripartal mammary gland. The phagocytic cells may be involved in the transportation of certain immunoglobulins into the neonate. In vitro colostral leukocytes exhibit a variety of immunological activities such as blastogenesis after mitogenic and antigenic stimulation, cytotoxicity and phagocytosis, but the medium milk confines these activities in comparison with those of blood leukocytes. Intact colostral leukocytes reach the gut of the gut of the newborn and may even cross the intestinal wall, gaining access to the neonates system and influencing its immunologic reactions, e.g. hypersensitivity and antibody-formation. The knowledge on the significance of colostral leukocytes for the protection against infection of the neonate is still limited.

The influence of colostral leukocytes on the immune system of the neonatal calf. IV. Effects on bactericidity, complement and interferon; synopsis.

The influence of colostral leukocytes on the bactericidity of whole blood of calves against a strain of E. coli and on the activities of haemolytic complement and interferon-alpha (the antiviral activity of sera resisting an acidic treatment at pH 2 for 6 h) in the serum was investigated during a period of 4 weeks using 4 experimental groups. The calves received either complete colostrum (COL+, n = 16), cell-depleted colostrum (COL-, n = 16), cell-supplemented milk substitute (MS+, n = 7) or pure milk substitute (MS-, n = 6) during their first three days of life. The bactericidity of whole blood of the COL+ group was significantly higher on the second and third days of life while the activity of haemolytic complement was lower after the first week as compared to the COL- group. No interferon-alpha was detectable in the sera of both COL groups. The bactericidity of the MS groups was significantly lower than that of the COL groups after the first day of life. It was significantly lower in the MS+ group after one week of life while the activity of haemolytic complement was higher than that of the MS- group. Three out of 5 MS- and only one out of 7 MS+ calves had low titres of interferon-alpha in their sera on the third day. Three out of 6 MS- calves died and 5 out of 7 MS+ animals. The mean day of death was 4.0 in the MS- and 8.4 in the MS+ group. Based on the in vitro results of this and the previous three communications it can be concluded that leukocytes which are an integral part of normal bovine colostrum, influence immunological reactions of the calf and that they may enhance its defence against infection. Colostral leukocytes in the absence of humoral components of the colostrum are not able to prevent fatal losses in the calves due to natural infection, although their influence on immune responses of the calves was detectable in vitro.

[Use of a Coffea arabica tosta extract for the prevention and therapy of polyfactorial infectious diseases in newborn calves]. / Einsatz eines Coffea arabica tosta Extrakts zur Prophylaxe und Therapie polyfaktorieller Infektionskrankheiten neugeborener Kälber.

Two studies have been carried out to evaluate the prophylactic and therapeutical effect of a 30%-extract from the coffee-bean seeds Coffeae arabicae on infectious diseases in newborn calves. 1. Within a large cattle-herd, which endemically showed a high proportion of infections within the gastroenteric and/or respiratory systems in calves, a randomised placebo-controlled double-blind study has been done. 50 newborn calves were given a subcutaneous injection of 10 ml Coffea-preparation 30% on first and third day of life. Another 50 calves received physiological saline as control. An index was set up which allowed to daily evaluate and compare body-temperature, consistency of feces, exsiccation-degree and breathing-rate of the animals. Besides this the number of therapeutical interventions and the number of days with disease-symptoms were recorded. Calves treated with Coffea-extract showed: on first and second day of life less animals with body-temperature below physiological values (p < 0.001 or 0.1 resp.), during the first period of diarrhea (between fourth and sixth day) significantly lower tendency of diarrhea (p < 0.1; 0.001; 0.005 resp.), after the second period of diarrhea (around the 9th day of life) a better and quicker recovery and a lower tendency of exsiccation (p < 0.05 on day 10 and 11) as the control-calves. Besides this the average duration of illness was shorter (4.7 instead of 7 days) and the average number of therapeutical interventions were less (3.1 instead of 4.5) than in control-calves. 2. Within four cattle-herds endemically showing a high rate of diarrhea in newborn calves the morbidity in a total of 371 animals could be dropped from about 45% to 10% by prophylactic administration of one to three s.-c.-injections of 10 ml Coffea-preparation together with one or two million i.U.Vit.A. one time perorally. For prophylactic use two injections of coffea preparation. on day 1 and 4 of life proved to be efficient under the given circumstances. Therapeutically the daily administration of a combination of Coffea-extract together with oral drugs containing tannic substances and diet-feed could reduce the mortality in animals with acute disease to about 30%.

Hospital-wide outbreak of Burkholderia contaminans caused by prefabricated moist washcloths.

We experienced a hospital outbreak of Burkholderia contaminans (Burkholderia cepacia Group K) in a German university hospital with two campuses. Cases were defined as the microbiological detection of B. cepacia complex (BCC) in any clinical specimen sent to the laboratory during 30 June to 21 October 2008. Species identification of BCC was performed by recA gene sequencing, followed by pulsed-field gel electrophoresis (PFGE; SpeI digest) for clonal identity. In total, 61 BCC-positive cases were diagnosed at the two campuses. At least nine patients contracted a ventilator-associated pneumonia with BCC. One patient suffered an infection of a pacing wire insertion site and four patients had septicaemia. Sixteen patients died in hospital, none thought to be due to the outbreak strain. BCC was eventually found in packages of moist prefabricated washcloths used for intensive care patients. German healthcare authorities were informed and a Europe-wide alarm (RAPEX) was initiated through the systems to prevent infections in other hospitals. PFGE proved clonal identity between isolates from clinical specimens and washcloths of both campuses. After elimination of the contaminated washcloths no further cases occurred. This example of a relatively newly introduced product raises the question of whether current regulations are adequate to protect consumers. For critically ill patients, care products should be carefully evaluated. In case of infections due to contaminated products, immediate communication to healthcare authorities is required, including RAPEX warning if products are sold across Europe.

Hepatitis viruses and the safety of blood donations.

Since the beginning of blood transfusions concomitant transmission of viral hepatitis has been a frequent and serious side-effect. A first measure to reduce the frequency of transmission was the screening of blood donors for elevated levels of liver enzymes in the blood, which was introduced in Germany in the 1960s, but not in most other countries. After the discovery of hepatitis B virus (HBV), donors in all countries have been screened since the 1970s for its surface antigen (HBsAg). When it was realized that there was at least one other type of virus that was even more frequently transmitted, screening for liver enzymes and HBV antibodies (anti-HBc) was introduced as a surrogate marker in most, but not all, countries in the 1980s. Furthermore, donors at risk for parenterally transmitted viruses were excluded. The discovery of the hepatitis C virus (HCV) genome and the development of sensitive anti-HCV assays has meant that reliable detection of persistently infected HCV carriers has been possible since 1991. Recently infected donors, however, are infectious for several weeks or months before anti-HCV is detectable. Therefore, starting in April 1999 all donations in Germany have to be tested, by nucleic acid amplification tests, for the presence of HCV RNA, although preliminary experience shows that such recent HCV infections are very rare. Newly detected viruses, named GBV-C or HGV and TTV, have been detected in patients with non-A-E post-transfusion hepatitis, but their association with the disease seems to be coincidental. These viruses cause persistent viraemia and are quite prevalent world-wide, but do not cause any known disease. At present, transfusion-transmitted hepatitis has been virtually eliminated, and any improvement in safety will be very small and will require huge costs.

Safety and quality criteria for autologous blood donations. Committee for Virus Safety of the Society for Virology.

The authors discuss the quality control criteria for autologous blood donations. They recommend that all autologous donations be tested for serological markers of blood-transmissible diseases. Marker-positive donors should not donate autologous blood. If autologous donation seems indispensable, infectious donations should be clearly marked as infectious.