Read the following article and answer the question USE YOUR OWN WORDS! Copying a
ID: 90528 • Letter: R
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Read the following article and answer the question
USE YOUR OWN WORDS! Copying any text from the article is forbidden (this includes nearly copying, changing only a few words)
1. The figures in this paper each correspond to the results from entirely different experiments, asking entirely different questions. For both figures, explain WHY the experiment was performed, and how the results answered the experimenters' question. – Note: I'm asking about the experiments performed, not the figures themselves. a) Figure 1: b) Figure 2:
Veterinary Paras itology 209 (2015) 281-284 Contents lists available at ScienceDirect Veterinary Parasitology ELSEVIER journal homepage: www.elsevier.com/locate/vetpar Short Communication Doxycycline levels and anti-Wolbachia antibodies in sera from dogs experimentally infected with Dirofilaria immitis and treated with a combination of ivermectin/doxycycline A. Menozzia, S. Bertinia, L. Turin b, P. Serventia, L Kramer C. Bazzocch Dip Scienze Medico-Veterinarie, Universita di Parma, via del Taglio 10. 43126Parma. Italy Dip di Scienze Veterinarie e Sanita Pubblica, Universita degli Studi di Milano, via Celoria 10, 20133 Milano, ltaly ABSTRACT ARTICLE I N FO Article history: Sera from Dirofilaria immitis-experimentally infected dogs treated with a combination of Received 5 January 2015 vermectin/doxycycline were analy for doxycycline levels by HPLC and anti-Wolbachia Received in revised form 19 February 2015 Surface Protein (rWSP) antibodies by ELISA and compared with sera from dogs treated Accepted 20 February 2015 with doxycycline alone. Results show that doxycycline levels were not statistically different between the two groups. Circulating anti-WSP antibody titres were markedly lower in both treatment groups when compared to control D immitis infected dogs, indicating that Dirofilaria immitis doxycycl e is able to reduce Wolbachia and prevent the immune response against the Doxycycline bacteria. The combination treatment protocol has been shown to be highly adulticidal and vermectin further studies are needed to better understand the interaction between doxycycline and ermectin in D immitis infected dogs. 2015 Elsevier BV. All rights reserved 1. Introduction has offered an interesting alternative for the treatment of HWD. Indeed, Wolbachia is necessary for the reproductive Heartworm infection (HW: Dirofilaria immitis) in dogs capacity and long-term survival of those filarial para causes chronic pulmonary disease that, if left untreated sites that harbour the endosymbiont.The adulticide effects can lead to right-side congestive heart failur e. Currently, of doxycycline (DOXY) have been studied in D. immitis the only registered drug for adul cide therapy in dogs experimentally infected dogs (Bazzocchi et al.. 2008). No with heartworm disease (HWD) is melarsomine dihy. significant adulticide effects at 8 months post infection fol mmiticide Merial) drochloride Due to concerns of lowing several cycles of DOXY was observed, even though severe, post-treatment thromboembolism in some dogs treatment was able to reduce Wolbachia populations. The er et al., 2008) and recent problems with availabil same study reported that when DOXY was combined with ty of melarsomine on several international markets, there the macrocyclic lactone ivermectin (IVM), adulticide effi is increasing interest in alternative adulticide treatments cacy was approximately 80%vs.9%when dogs were treated (Colby et al., 2011) with DOXY alone. The adulticide effect of this combina The recent targeting ofthe bacterial endosymbiont Wol tion has also been confirmed in naturally infected dogs bachia, through antibiotic therapy of the infected host. (Grandi et al 2010) It is not clear why the two drugs work better together in eliminating a large population of heart worms in a relatively short period of time (8-10 months). t is not yet known if this is due to a simple summation Corresponding author Tel: 39 0521 032715: fax:+39052103270. effect or ifthere exists a certain synergism between the two E-mail address kramerlheunipr it (L Kramer http://dx doi.org/10.1016/jvetpar.2015.02.023 0304-4017jo 2015 Elsevier BV.All rights reserved.Explanation / Answer
Figure 1
The subjects used in this analytical research are Dirofilaria immitis infected dogs and the sample
being analysed is dogs sera. The infected dogs are treated with a combination drug of ivermectin/ doxycycline. The experimental approach designed is HPLC which stands for high perfromance liquid chromatography. As the researcher wants to analyse only doxycycline levels from a combination drug, HPLC experiment was
performed.This chromatographic technique seperates mixture of components( ivermectin/ doxycycline) into an individual component (doxycycline)and also quantifies it. The set of conditions suitable for separation of the mixture are established using stationary phase, mobile phase, solvents and buffers. The results are obtained at different time points. The sera from infected dogs which are not receiving any treatment and sera from infected dogs which are treated with doxycline alone ,are also run through HPLC column. The results collected from all are compared for analysis.
The interpretation of chromatographic results by this method was difficult because there is a wide range of variability in serum concentrations among dogs versus timepoints.
Figure 2
ELISA enzyme linked immnunosorbent assay is an analytical tool that detects and measures antibodies in a blood sample. It helps in determination of antibodies related to certain infections. Therefore, to determine the presence of anti-wolbachia surface protein (rWSP) antibodies in dogs infected with Dirofilaria immitis , the researcher performed ELISA experiment.
In this experiement anti dog IgG HRP conjuagated antibody is made in response to infection. The samples
containing sera of dogs receiving no treatment,treated with combination drug and treated with doxycycline alone are run and results are collected at different time points.
The analysis showed that the dogs receiving combination drug and doxycycline alone have markedly lower values for anti-WSP antibodies when compared to the untreated infected dogs sample.
The results from these two experiments answered experiementers question as follows ,the adulticide effect from
combination drug is not due to higher drug concentration of DOXY nor due to lack of efficacy in the removal of wolbachia from worm tissues.
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