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Introduction: In addition to hand washing and wearing masks, social distancing and reducing exposure time to <15 minutes are the most effective measures against the spread of COVID-19. Unfortunately, three of these guidelines are very difficult, if not impossible, for nursing babies: they cannot wear masks, stay six feet away from the lactating breasts, nor consistently finish within 15 minutes while nursing. We report a case of a nursing mother with SARS-CoV-2 infection, documenting changes of immune cells and cytokines in breast milk with and without the infection. Case Description: With Institutional Review Board (IRB) approval, we obtained expressed breast milk samples from a lactating mother before and during SARS-CoV-2 infection as documented by reverse transcription-PCR. Using flow cytometry analysis, we measured the immune cell profiles and expression of cytokines such as interferon alpha (IFNα) in milk leukocytes before and during infection. Results: There was an eightfold increase in IFNα+ milk leukocytes, from 1% before SARS-CoV-2 infection to 8% when actively infected. The milk macrophages showed the highest increase in IFNα expression. Both T and B lymphocytes showed mild increase. Innate lymphoid cells, neutrophils, and natural killer cells showed no increase in IFNα expression and the dendritic cells actually showed a reduction. Conclusion: We document the presence and high expression of IFNα in the breast milk macrophages of a lactating mother with confirmed COVID-19, compared with her milk before the infection.

In addition to providing life-giving nutrients and other substances to the breastfed infant, human milk can also represent a vehicle of pathogen transfer. As such, when an infectious disease outbreak, epidemic, or pandemic occurs-particularly when it is associated with a novel pathogen-the question will naturally arise as to whether the pathogen can be transmitted through breastfeeding. Until high-quality data are generated to answer this question, abandonment of breastfeeding due to uncertainty can result. The COVID-19 pandemic, which was in full swing at the time this document was written, is an excellent example of this scenario. During these times of uncertainty, it is critical for investigators conducting research to assess the possible transmission of pathogens through milk, whether by transfer through the mammary gland or contamination from respiratory droplets, skin, breast pumps, and milk containers, and/or close contact between mother and infant. To promote the most rigorous science, it is critical to outline optimal methods for milk collection, handling, storage, and analysis in these situations, and investigators should openly share their methods in published materials. Otherwise, the risks of inconsistent test results from preanalytical and analytical variation, false positives, and false negatives are unacceptably high and the ability to provide public health guidance poor. In this study, we provide "best practices" for collecting human milk samples for COVID-19 research with the intention that this will also be a useful guide for future pandemics.

Aim: The objective of our study was to determine whether the SARS-CoV-2-positive mothers transmit the virus to their hand-expressed colostrum. Methods: This is an observational prospective study that included pregnant women who tested positive for SARS-CoV-2 by PCR test on a nasopharyngeal swab at the moment of childbirth and who wanted to breastfeed their newborns. A colostrum sample was obtained from the mothers by manual self-extraction. To collect the samples, the mothers wore surgical masks, washed their hands with an 85% alcohol-based gel, and washed their breast with gauze that was saturated with soap and water. Results: We obtained seven colostrum samples from different mothers in the first hours postdelivery. SARS-CoV-2 was not detected in any of the colostrum samples obtained in our study. Conclusion: In our study, breast milk was not a source of SARS-CoV-2 transmission. Hand expression (assuring that a mask is used and that appropriate hygienic measures are used for the hands and the breast), when direct breastfeeding is not possible, appears to be a safe way of feeding newborns of mothers with COVID-19.

Background: Limited data are available on the perinatal and postnatal transmission of novel coronavirus disease 2019 (COVID-19). The Centers for Disease Control and Prevention (CDC) and World Health Organization (WHO) recommended breastfeeding with necessary precautions to mothers with COVID-19. Case Presentation: A 20-year-old pregnant woman with no symptoms of COVID-19 presented to the hospital for delivery at 39 weeks of gestation. She was tested for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) by reverse transcriptase polymerase chain reaction (RT-PCR) because her father had been diagnosed with COVID-19. A nasopharyngeal swab RT-PCR test was positive for SARS-CoV-2. Therefore, the baby and the mother were cared for separately after delivery. Breast milk obtained after first lactation was tested by real-time RT-PCR and was positive for SARS-CoV-2. Conclusions: In this article, we aimed to report the presence of SARS-CoV-2 in breast milk. Although further studies are needed, this situation may have an impact on breastfeeding recommendations.

Background: The aim of this study was to evaluate the freeze-thawing method in reducing viral load of cytomegalovirus (CMV) in breast milk of mothers of preterm infants., Materials and Methods: In this study, 169 nursing mothers of preterm infants were evaluated serologically for CMV infection. We found 29 (17.15%) nursing mothers who had serologic evidence of CMV infections consistent with recent infection (N = 2, IgG + IgM +) or prior infection (N = 27 IgG + IgM-). Two to 6 weeks after delivery, breast milk of seropositive mothers (N = 29) was evaluated for CMV load by real-time polymerase chain reaction (PCR) before and after freezing at -20°C for 72 hours. All preterm infants with milk results positive for DNA particles of CMV (N = 25) were followed up for signs and symptoms of CMV infection until 3 months of age, together with urine testing for CMV by real-time PCR., Results: Examination of breast milk by real-time PCR of the seropositive mothers showed 25 of them with CMV DNA particles in their breast milk. After freezing and thawing, we found virus DNA in 4 of 25 (16%) of the breast milk samples and there was no CMV DNA particles in 21 (84%) of them. In these samples, mean viral load before freezing was 76.04 ± 34.08 copies/μL (20-135.00 copies/μL) and after freezing it was 6.75 ± 4.34 copies/μL (0.00-13.00 copies/μL). Freezing showed a significant decrease in viral load of the samples (p = 0.001)., Conclusion: The study showed that the freeze-thawing method is an effective method in reducing the CMV load in breast milk samples.

Introduction: A key strategy to prevent mother-to-child transmission of the human immunodeficiency virus (HIV) and to reduce infant morbidity and mortality includes providing the HIV-exposed premature infant with breast milk accompanied by dual anti-retroviral therapy (ART). The effects of HIV and ART on premature breast milk composition are largely unknown. The aim of the study was to assess and compare the breast milk composition of HIV-infected mothers receiving ART and HIV-uninfected mothers who gave birth to premature infants., Materials and Methods: Lactating HIV-infected women receiving ART (n = 38) and HIV-uninfected women (n = 36) with premature infants provided two breast milk samples on days 7 and 9, respectively, of lactation. Breast milk samples were analyzed for total energy, protein, carbohydrates, fat, phosphate, iron, zinc, and copper content., Results: Breast milk of HIV-infected women contained higher protein (1.95 versus 1.78 g/100 g; p = 0.04), fat (4.42 versus 3.49 g/100 g; p = 0.01), and copper (0.64 versus 0.56 mg/L; p = 0.02) levels; whereas carbohydrate (5.37 versus 6.67 g/100 g; p = 0.002) and zinc (5.26 versus 5.78 mg/L; p = 0.04) levels were lower compared with those of HIV-uninfected women. Zinc levels were significantly lower in HIV-infected women with early gestation periods, and the lowest levels were observed in women who received ART for ≤4 weeks (0.58 mg/L; p = 0.03). Total energy (78.22 versus 61.48 kCal/100 mL) and fat levels (5.39 versus 3.00 g/100 mL) were significantly higher in the late gestation period HIV-infected women. Copper levels (0.61 mg/L) were higher in the late gestation period women who received >4 weeks of ART exposure (p = 0.05)., Conclusion: Differences existed in the breast milk composition of HIV-infected women on ART compared with HIV-uninfected women. ART exposure period may influence breast milk composition.

Background: Cytomegalovirus (CMV)-infected human milk (HM) can lead to significant CMV morbidity and mortality in preterm very-low-birth weight infants. The eradication of CMV in HM while preserving its properties poses a major clinical challenge., Objective: We aimed to compare two methods used to neutralize the virus in HM, one recognized as partially effective (freezing) and another not tested to date (microwave exposure)., Materials and Methods: We sampled HM from 31 CMV-seropositive mothers whose infants were hospitalized at the Lis Maternity Hospital. Fifteen samples that were positive for CMV antigen were divided into five 5 mL aliquots: the first a control, the second was frozen at -20°C for 1 day, the third was frozen at -200°C for 3 days, and the fourth and fifth aliquots were exposed for 30 seconds to microwave radiation at a low-power setting (500 W) and high-power setting (750 W), respectively., Results: Only microwave radiation at a high-power setting led to complete neutralization of CMV in all samples. Low-power microwave irradiation had a 13% failure rate while 3-day freezing and 1-day freezing had failure rates of 7% and 20%, respectively., Conclusion: It is possible to eradicate CMV successfully in HM by using microwave radiation at a high-power setting. Further studies are needed to evaluate the effect of microwave heating on breast milk properties.

Breastmilk specimens from three women with acute hepatitis A virus (HAV) infection were studied. Anti-HAV immunoglobulin M and immunoglobulin G antibodies were detected in serum and breastmilk specimens of the three women. The three women also had serum HAV RNA. However, HAV RNA was detected only in two of the three breastmilk specimens. It is interesting that none of the three infants contracted clinical HAV infection. Furthermore, mothers with HAV infection should not be encouraged to discontinue breastfeeding.

Background: Human immunodeficiency virus type 1 (HIV-1) transmission through breastmilk is the chief modality through which HIV-1 is transmitted from HIV-1-infected mothers to their babies in developing countries, where alternative feeding options lack practical feasibility. The development of an approach to inactivate the HIV-1 virions ingested by an infant on a daily basis through breastmilk is thus of critical importance., Methods: Copper has potent virucidal properties. Stoichiometric concentrations of copper ions inactivate the HIV-1 protease, which is essential for viral replication. Cell-free and cell-associated HIV-1 infectivity is inhibited when the virus is exposed to copper oxide in a dose-dependent manner. Passage of high titers of a wide range of HIV-1 isolates, spiked in culture medium, through filters containing copper oxide powder resulted in their deactivation., Results: In the current study, we demonstrate that the infectivity of three different HIV-1 isolates, spiked in breastmilk obtained from HIV-1-seronegative donors, or of wild-type isolates found in breastmilk obtained from HIV-1-seropositive donors, is drastically reduced (>98%) when exposed to copper oxide., Conclusions: This study is proof of concept that copper oxide is efficacious against HIV-1 found in breastmilk and serves as the basis for further research aimed at determining the possible effects that copper may have on the nutritional and anti-infective properties of breastmilk. Furthermore, this supports the continuing study of the feasibility of developing a filtering device, such as an "at-the-breast" disposable shield that can be used discreetly and safely by HIV-1-infected mothers during breastfeeding.

Background and Objectives: The World Health Organization recommends human immunodeficiency virus (HIV)-positive mothers in resource-poor regions heat-treat expressed breastmilk during periods of increased maternal-to-child transmission risk. Flash-heat, a "low tech" pasteurization method, inactivates HIV, but effects on milk protein bioactivity are unknown. The objectives were to measure flash-heat's effect on antimicrobial properties of lactoferrin, lysozyme, and whole milk and on the digestive resistance of lactoferrin and lysozyme., Methods: Flash-heated and unheated breastmilk aliquots from HIV-positive mothers in South Africa were "spiked" with Staphylococcus aureus and Escherichia coli and then cultured for 0, 3, and 6 hours. Lysozyme and lactoferrin activities were determined by lysis of Micrococcus luteus cells and inhibition of enteropathogenic E. coli, respectively, measured spectrophotometrically. Percentages of proteins surviving in vitro digestion, lactoferrin and lysozyme activity, and bacteriostatic activity of whole milk in heated versus unheated samples were compared., Results: There was no difference in rate of growth of E. coli or S. aureus in flash-heated versus unheated whole milk (p = 0.61 and p = 0.96, respectively). Mean (95% confidence interval) antibacterial activity of lactoferrin was diminished 11.1% (7.8%, 14.3%) and that of lysozyme by up to 56.6% (47.1%, 64.5%) by flash-heat. Digestion of lysozyme was unaffected (p = 0.12), but 25.4% less lactoferrin survived digestion (p < 0.0001)., Conclusions: In summary, flash-heat resulted in minimally decreased lactoferrin and moderately decreased lysozyme bioactivity, but bacteriostatic activity of whole milk against representative bacteria was unaffected. This suggests flash-heated breastmilk likely has a similar profile of resistance to bacterial contamination as that of unheated milk. Clinical significance of the decreased bioactivity should be tested in clinical trials.

Background: Human breastmilk provides a rich source of commensal lactic acid bacteria (LAB) to the infant during breastfeeding and stimulates abundant growth and colonization of these bacteria at mucosal surfaces in the infant gastrointestinal tract. While conferring critical nutritional and immunologic support to the developing newborn, breastmilk also serves as a vehicle for human immunodeficiency virus type 1 (HIV-1) transmission from mother to child during breastfeeding. Whether breastmilk LAB confer protection against mucosal exposure to HIV-1 in breastfeeding infants is unknown., Study Design: In the present study, we sought to evaluate LAB isolated from the breastmilk of healthy women for the ability to inhibit HIV-1 infection in vitro. A total of 38 strains of breastmilk bacteria were evaluated in this study. Both heat-killed bacteria and cell-free conditioned supernatants from bacterial cultures were tested for the ability to inhibit infection with HIV-1 using viral isolates with tropism for CCR5 (R5), CXCR4 (X4), or R5/X4 dual-tropism., Results: Significant inhibition of R5-tropic HIV-1 was demonstrated using heat-killed bacteria, most notably among breastmilk strains of Lactobacillus and Pediococcus. Selected strains of breastmilk LAB also demonstrated significant inhibition of HIV-1 infection against virus with tropism for X4 and R5/X4., Conclusion: These results demonstrate for the first time that commensal LAB from human breastmilk inhibit HIV-1 infection in vitro and suggest a possible role for these bacteria in mucosal protection against HIV-1 in the breastfeeding infant.

Background: In Sweden preterm infants born <32 gestational weeks are fed maternal breastmilk or, if not available, donor breastmilk. Nutritional and immunological composition of human milk is affected by processing and storage procedures. Additionally, freezing of breastmilk may reduce cytomegalovirus transmission. The present recommendations for human milk use in Sweden are outdated. However, new guidelines to standardize routines are underway. This study was designed to document current routines pertaining to breastmilk use for preterm infants in Sweden., Methods: A questionnaire regarding breastmilk handling and routines was sent to all 36 neonatal units in Sweden in November 2006 and February 2007., Results: Of the 36 participating neonatal units 27 had their own milk bank. Milk donors were screened for human immunodeficiency virus, human T-lymphotropic virus, and hepatitis B and C viruses by 27, 14, and 22 of the milk banks, respectively. Bacterial culture was performed on donor milk in 24 milk banks. Donor milk was pasteurized in 22 milk banks. In 11 of the 36 neonatal units maternal milk was frozen to reduce the risk of cytomegalovirus transmission. No neonatal unit performed bacterial culture or pasteurization of maternal milk. Breast milk was kept frozen for a maximum of 3-6 months before use. Nutritional analysis of donor and/or maternal milk was performed in 25 units. All neonatal units enriched donor milk and maternal milk., Conclusions: Routines for breastmilk handling differ among the 36 neonatal units in Sweden. New guidelines can standardize the handling of human milk, thereby improving nutrition and minimizing the risk of breastmilk-induced infection in the preterm infant.

In the United States, concerns over the transmission of infectious diseases have led to donor human milk generally being subjected to pasteurization prior to distribution and use. The standard method used by North American milk banks is Holder pasteurization (63 degrees C for 30 minutes). The authors undertook an experiment to validate the effects of a high-temperature short-time (HTST) pasteurization process (72 degrees C for 16 seconds) on the bioburden of human milk. It was concluded that HTST is effective in the elimination of bacteria as well as of certain important pathogenic viruses.

Human cytomegalovirus (HCMV) can be transmitted through breast milk to neonates. Although healthy full-term infants rarely develop symptoms of CMV infection; premature or low-birth-weight infants can experience symptomatic infection that is occasionally severe. There is limited information on the long-term effects of postnatal CMV infection in premature infants, suggesting that these infants do not develop cognitive function delays or hearing loss, although those with intrapartum infection do. Readily available methods of treating breast milk to inactivate the CMV either diminish the immunologic and nutritive benefits of breast milk or incompletely inactivate the virus. This review considers the data on measuring CMV in breast milk, the recent clinical studies on CMV transmission via breast milk, reported methods of inactivation of CMV in breast milk, and immunologic factors that may play a role in transmission. CMV-IVIG treatment needs further evaluation but appears promising. Recommendations are made to help address the issue of CMV transmission to premature infants in clinical practice in the neonatal intensive care unit (NICU).