Integrating Blood Collection Within Household Surveys: Lessons Learned From Nesting a Measles and Rubella Serological Survey Within a Post-Campaign Coverage Evaluation Survey in Southern Province, Zambia

Age-specific population immunity to many vaccine-preventable diseases can be measured using serological surveys. However, stand-alone serological surveys are infrequently conducted in low- and middle-income countries because of costs, operational challenges, and potential high refusal rates for blood collection. Nesting a serosurvey within a household cluster survey may overcome some of these challenges. We share lessons learned from nesting a serosurvey within a measles and rubella vaccination post-campaign coverage evaluation survey (PCES). In 15 of the 26 PCES clusters in Southern Province, Zambia, we collected dried blood spots from 581 participants aged 9 months and older. Household participation rates for the main PCES were higher in the serosurvey clusters (86%) than PCES-only clusters (71%), suggesting that a serosurvey can be successfully integrated without adversely affecting PCES participation. Among households that participated in the PCES, 80% also participated in the serosurvey and 86% of individuals available in the household provided a blood sample for the serosurvey. Substantial planning and coordination, additional staff training, and community mobilization were critical to the success of the serosurvey. Most challenges stemmed from using different data collecting tools and teams for the serosurvey and PCES. A more efficient design would be to fully integrate the serosurvey by adding blood collection and additional questions to the PCES

Role of CpSUB1, a subtilisin-like protease, in Cryptosporidium parvum infection in vitro.

The apicomplexan parasite Cryptosporidium is a significant cause of diarrheal disease worldwide. Previously, we reported that a Cryptosporidium parvum subtilisin-like serine protease activity with furin-type specificity cleaves gp40/15, a glycoprotein that is proteolytically processed into gp40 and gp15, which are implicated in mediating infection of host cells. Neither the enzyme(s) responsible for the protease activity in C. parvum lysates nor those that process gp40/15 are known. There are no furin or other proprotein convertase genes in the C. parvum genome. However, a gene encoding CpSUB1, a subtilisin-like serine protease, is present. In this study, we cloned the CpSUB1 genomic sequence and expressed and purified the recombinant prodomain. Reverse transcriptase PCR analysis of RNA from C. parvum-infected HCT-8 cells revealed that CpSUB1 is expressed throughout infection in vitro. In immunoblots, antiserum to the recombinant CpSUB1 prodomain revealed two major bands, of approximately 64 kDa and approximately 48 kDa, for C. parvum lysates and proteins "shed" during excystation. In immunofluorescence assays, the antiserum reacted with the apical region of sporozoites and merozoites. The recombinant prodomain inhibited protease activity and processing of recombinant gp40/15 by C. parvum lysates but not by furin. Since prodomains are often selective inhibitors of their cognate enzymes, these results suggest that CpSUB1 may be a likely candidate for the protease activity in C. parvum and for processing of gp40/15. Importantly, the recombinant prodomain inhibited C. parvum infection of HCT-8 cells. These studies indicate that CpSUB1 plays a significant role in infection of host cells by the parasite and suggest that this enzyme may serve as a target for intervention

Measles and Rubella Serosurvey Identifies Rubella Immunity Gap in Young Adults of Childbearing Age in Zambia: The Added Value of Nesting a Serological Survey Within a Post-Campaign Coverage Evaluation Survey

Background: Serological surveys can potentially complement vaccine coverage surveys, such as post-vaccination campaign coverage evaluation surveys (PCES), by providing direct information on population immunity within and outside the target age range of the mass vaccination campaign. We estimate age-specific population immunity to measles and rubella viruses in Southern Province, Zambia, and assess the value of adding serological data to vaccination coverage estimates by nesting a serological survey within a PCES. Methods: Dried blood spots (DBS) from fingerprick blood were collected from all individuals ages nine months or older in households participating in the PCES and tested for measles and rubella virus-specific immunoglobulin G (IgG) by enzyme immunoassay (Siemens Enzygnost, Marburg, Germany). Results: Overall seroprevalence was 95.5% (95% CI: 92.8, 97.2) for measles virus-specific IgG and 97.7% (95% CI: 96.0, 98.7) for rubella virus-specific IgG. Rubella seroprevalence was 98.4% (95% CI: 95.9, 99.4) among children eligible for the MR vaccination campaign, significantly higher than the reported measles-rubella (MR) vaccination campaign coverage of 89.8% (p = 0.003), and higher than the 91.3% rubella seroprevalence for adolescents and adults 16–30 years of age (p = 0.049). Conclusion: Seroprevalence to measles and rubella viruses in children younger than 16 years of age was significantly higher than expected from vaccination coverage estimates, likely reflecting exposure to wild-type viruses and underreporting of vaccination. The serosurvey revealed rubella immunity gaps among women 16–30 years of age, precisely the age group in which protection from rubella is most important to prevent congenital rubella syndrome. Nesting serological surveys within existing surveys can leverage resources and infrastructure while providing complementary information important to immunization programs. © 2019 The Author(s

Cryptosporidium

Cryptosporidium sp. infects the gastrointestinal tract of a wide range of vertebrates, including domestic and livestock animals as well as humans. Cryptosporidiosis of neonatal farm ruminants causes considerable economic losses as the disease is commonly associated with intense diarrhea leading to an impaired growth, a decreased performance and production, and often animal death. The highly infective oocyst stage is excreted with the feces and disseminated into the environment, contaminating water and food. Neonatal calves are a major reservoir of the zoonotic C. parvum, which causes, in addition to the anthroponotic C. hominis, human cryptosporidiosis and is of considerable public health concern. Currently, no vaccine or efficient drug is available against the disease. From a veterinarian economical point of view, C. parvum is the most important species among the 30 recognized species infecting bovines, lamb, goats, pigs, horses, and dogs worldwide. After the discovery of C. parvum by Tyzzer in the year 1912, the taxon Cryptosporidium has been classified into coccidia. However, recent findings on the Cryptosporidium life cycle and molecular phylogenetic evidence resulted in the reclassification of Cryptosporidium into the gregarines. This novel placement appreciates previously underestimated and/or neglected features of Cryptosporidium that are common to gregarines such as a low host specificity and/or the possibility to survive without a host.Fil: Tomazic, Mariela Luján. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; ArgentinaFil: Garro, Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; ArgentinaFil: Schnittger, Leonhard. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina. Universidad de Morón; Argentin